Methods of preparing pharmaceutical solid state forms

ABSTRACT

The invention provides and describes solid state 17α-ethynyl-5α-androstane-3α,17β-diol including amorphous and crystalline forms and specific polymorphic forms thereof. Anhydrates and solvates of 17α-ethynyl-5α-androstane-3α,17β-diol include Form III anhydrate and Form I solvate. The invention further relates to solid and suspension formulations containing 17α-ethynyl-5α-androstane-3α,17β-diol in a described solid state form and use of the formulations to treat cancers or precancers such as prostate cancer or breast cancer in subjects or human patients. The invention also relates to methods to make liquid formulations from solid state forms of 17α-ethynyl-5α-androstane-3α,17β-diol and uses of such formulations in treating the described conditions.

CROSS REFERENCE TO RELATED APPLICATIONS

This nonprovisional U.S. patent application is a divisional of andclaims priority under 35 U.S.C. §121 to U.S. nonprovisional applicationSer. No. 12/370,510 filed Feb. 12, 2009, now U.S. Pat. No. 8,518,922,which is a continuation of and claims priority under 35 U.S.C. §365(c)to PCT international patent application serial No. PCT/US09/33280 filedFeb. 5, 2009, which claims priority under 35 U.S.C. §119(e) from U.S.provisional application Ser. No. 61/093,694 filed Sep. 2, 2008, and U.S.provisional application Ser. No. 61/026,472, filed Feb. 5, 2008, all ofwhich are incorporated herein by reference in their entireties.

FIELD OF THE INVENTION

The invention relates to17-ethynyl-10R,13S-dimethyl-2,3,4,5S,6,7,8R,9S,10,11,12,13,14S,15,16,17-hexadecahydro-1H-cyclopenta[a]-phenanthrene-3S,17S-dioland its solid state forms, including crystalline, polymorph,pseudopolymorph and amorphous forms and methods for their preparing thesolid state forms. The invention further relates to solid formulationscomprising the solid state forms and to methods for using the solidstate forms, including the polymorph forms and pseudopolymorph forms, inpreparing solid and liquid formulations and uses of these formulationsfor the treatment of cancer, including hormone sensitive or hormoneassociated cancers such as breast cancer, prostate cancer, and for thetreatment of precancers and hyperplasias such as benign prostatehyperplasia. Unit dosage forms for the solid and liquid formulations arealso included.

BACKGROUND OF THE INVENTION

The ability of a substance to exist in more than one crystalline form isgenerally referred to as polymorphism and these different crystallineforms are usually named “polymorphs” and may be referred to by certainanalytical properties such their X-ray powder diffraction (XRPD)patterns. In general, polymorphism reflects the ability of a molecule tochange its conformation or to form different intermolecular andintramolecular interactions. This can result in different atomarrangements that is reflected in the crystal lattices of differentpolymorphs. However, polymorphism is not a universal feature of solids,since some molecules can exist in one or more crystal forms while othermolecules cannot. Therefore, the existence or extent of polymorphism fora given compound is unpredictable.

The different polymorphs of a substance posses different crystal latticeenergies and thus each polymorph typically shows one or more differentphysical properties in the solid state, such as density, melting point,color, stability, dissolution rate, flowability, compatibility withmilling, granulation and compacting and/or uniformity of distribution[See, e.g., P. DiMartino, et al., J. Thermal Anal. 48:447-458 (1997)].The capacity of any given compound to occur in one or more crystallineforms (i.e. polymorphs) is unpredictable as are the physical propertiesof any single crystalline form. The physical properties of a polymorphicform may affect its suitability in pharmaceutical formulations. Thoseproperties can affect the stability, dissolution and bioavailability ofa solid-state formulation, which subsequently affects suitability orefficacy of such formulations in treating disease.

An individual polymorph having one or more desirable properties can besuitable for the development of a pharmaceutical formulation havingdesired property(ies). Existence of a compound with a polymorphicform(s) having undesirable properties can impede or prevent developmentof the polymorphic form as a pharmaceutical agent.

In the case of a chemical substance that exists in more than onepolymorphic form, the less thermodynamically stable forms canoccasionally convert to the more thermodynamically stable form at agiven temperature after a sufficient period of time. When thistransformation is not rapid, such a thermodynamically unstable form isreferred to as a “metastable” form. In some instances, the stable formexhibits the highest melting point, the lowest solubility, and themaximum chemical stability. In other cases, the metastable form mayexhibit sufficient chemical and physical stability under normal storageconditions to permit its use in a commercial form. In this case, themetastable form, although less thermodynamically stable, may exhibitproperties desirable over those of the stable form, such as enhancedsolubility or better oral bioavailability. Likewise, the amorphous formof an active pharmaceutical ingredient may have different solubility incomparison to a given crystalline material due reduction of crystallattice forces in the amorphous material that must be overcome to effectdissolution in aqueous or non-aqueous liquids.

SUMMARY OF THE INVENTION

In a principal embodiment, the invention provides new solid state formsof17-Ethynyl-10R,13S-dimethyl-2,3,4,5S,6,7,8R,9S,10,11,12,13,14S,15,16,17-hexadecahydro-1H-cyclopenta[a]phenanthrene-3S,17S-diol,which is represented by Formula 1. This compound is suitable fortreating a hyperproliferation condition such as cancer or precancer andin particular, a hormone sensitive or associated cancer, precancer orbenign hyperplasia, such as prostate cancer, breast cancer, prostaticintraepithelial neoplasia or benign prostatic hypertrophy.

The compound of Formula 1 (hereafter also referred to as Compound 1 or17α-ethynyl-5α-androstane-3α,17β-diol) has been prepared in amorphousand crystalline forms, and in particular, crystalline forms referredherein as Form I, Form III, Form IV, Form V, Form VI, Form VII and FormVIII.

One embodiment of the invention is directed to a particular crystallineform of Compound 1 (e.g., Form I, Form III, Form IV, Form V, Form VI,Form VII, Form VIII) substantially free or essentially free of othercrystalline or amorphous forms of Compound 1.

In certain embodiments, the present invention is directed to aparticular polymorph form (e.g., Form III) or pseudopolymorph form(e.g., Form I) of Compound 1 that is substantially free or essentiallyfree of other polymorph or pseudopolymorph forms of Compound 1.

Another embodiment of the invention is directed to amorphous Compound 1,typically wherein the amorphous material is substantially free oressentially free of crystalline Compound 1.

In certain embodiments, the present invention provides methods ofmaking, isolating and/or characterizing the solid state forms of theinvention. Some of these embodiments are directed to methods to prepareCompound 1 in crystalline form. Other such embodiments are directed tomethods to prepare Compound 1 in amorphous form.

In some embodiments a solid state form of Compound 1 is characterized oridentified by methods comprising X-ray Powder Diffraction (XRPD) and oneor more thermal methods including Differential Thermal Analysis (DTA),Differential Scanning calorimetry (DSC), Thermogravimetric Analysis(TGA) and melting point measurements.

In some embodiments a solid state form of Compound 1 is characterized oridentified by methods including XRPD and a vibrational spectroscopymethod such as Raman spectroscopy.

In some embodiments a solid state form of Compound 1 is characterized oridentified by methods including single crystal X-ray diffraction.

In some embodiments a solid state form of Compound 1 is characterized oridentified by methods including ¹H-NMR, elemental analysis, Karl-Fishertitration, thermogravimetric analysis or a combination thereof.

In some embodiments a crystalline form of Compound 1 is identified orcharacterized by a method comprising the steps of (1) obtaining unitcell parameters for a reference crystalline form; (2) obtaining one ormore high resolution XRPD patterns for the crystalline form to beidentified or characterized and indexing the high resolution XRPDpattern so obtained; (3) determining unit cell parameters for thecrystalline form to be identified or characterized from the indexed highresolution XRPD pattern(s); and (4) comparing the unit cell parametersfor the reference crystalline form and the crystalline form to beidentified. In certain embodiments of this method, the unit cellparameters for the reference crystalline form is obtained from singlecrystal X-ray data. In other embodiments of this method the unit cellparameters for the reference crystalline form are obtained from indexedhigh resolution XRPD pattern(s). In still other embodiments of thismethod the crystalline form to be identified or characterized is apolymorph or pseudopolymorph and the reference is another polymorph orpseudopolymorph to be identified or characterized. In some of thesemethods the crystalline form to be identified is a pseudopolymorph andthe reference is an anhydrate, preferably an isostructural anhydrate. Instill other embodiments of these methods, the unit cell parameters thatare compared are crystal volumes derived for an isostructural referencecrystalline form and the crystalline form to be characterized.

Other embodiments of the invention are directed to solid formulationscomprising a solid state form of Compound 1 and methods for preparationof the solid formulation.

In certain embodiments, the present invention encompasses the use of thesolid state forms of the invention as a final drug product.

Other embodiments of the invention are directed to pharmaceuticallyacceptable formulations comprising a particular crystalline form (e.g.Form I, Form III, Form IV, Form V, Form VI, Form VII, Form VIII) ofCompound 1 that is substantially free or essentially free of other solidstate forms, such as amorphous or other crystalline forms of Compound 1,and methods for preparation of the formulations.

Still other embodiments of the invention are directed to liquidformulations prepared by contacting or admixing at least one solid stateform of Compound 1 with a liquid excipient into which Compound 1 hassufficient solubility, optionally in the presence of another excipient,and methods for preparation of the liquid formulation.

Yet another embodiment of the invention is directed to methods fortreating a cancer or hyperproliferation condition such as a hormoneassociated cancer or hormone sensitive cancer including ovarian cancer,endometrial cancer, prostate cancer or breast cancer in a subject with asolid formulation comprising a solid state form of Compound 1 such asamorphous or a crystalline form of Compound 1.

Yet another embodiment of the invention is directed to methods fortreating a hormone associated cancer or hormone sensitive cancer such asovarian cancer, endometrial cancer, prostate cancer or breast cancer ina subject with a solid formulation comprising a particular crystallineform (e.g. Form I, Form III, Form IV, Form V, Form VI, Form VII, FormVIII) of Compound 1 that is substantially free of other solid stateforms, such as amorphous and other crystalline forms, of Compound 1

Another embodiment of the invention is directed to methods for treatingendometriosis or benign prostate hyperplasia in a subject with a solidformulation comprising a solid state form of Compound 1 such asamorphous or a crystalline form of Compound 1.

Invention embodiments also include the use or Compound 1 in amorphous orcrystalline form for the preparation of a medicament for the treatmentor prophylaxis of a hormone associated or sensitive cancer, precancer orhyperplasia such as prostate cancer, breast cancer, prostaticintraepithelial neoplasia or benign prostatic hypertrophy.

Still other embodiments are directed to methods for preparing liquidformulations using a solid state form of Compound 1 and uses of suchformulations for treating a hormone associated cancer or hormonesensitive cancer.

Other embodiments and advantages of the present invention are asdescribed elsewhere in the specification including the numberedembodiments and the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 provides a low resolution X-ray powder diffraction pattern of asynthesis product obtained in the preparation of bulk Compound 1.

FIG. 2 provides a high resolution X-ray powder diffraction pattern of asample comprising Form III 17α-ethynyl-5α-androstane-3α,17β-diol.

FIG. 3 provides differential scanning calorimetry and thermogravimetricanalysis thermograms of a sample comprising Form III17α-ethynyl-5α-androstane-3α,17β-diol.

FIG. 4 provides a proton NMR spectroscopy spectrum of a samplecomprising Form III 17α-ethynyl-5α-androstane-3α,17β-diol.

FIG. 5 provides an expanded view in a region of the NMR spectroscopyspectrum of FIG. 4.

FIG. 6 provides a Raman spectroscopy spectrum for a sample comprisingForm III 17α-ethynyl-5α-androstane-3α,17β-diol.

FIG. 7 provides an expanded view in a region of the Raman spectroscopyspectrum of FIG. 6.

FIG. 8 provides a microscope photograph of crystals of Form III17α-ethynyl-5α-androstane-3α,17β-diol under 10× magnification.

FIG. 9 provides a high resolution X-ray powder diffraction pattern of asample comprising Form I 17α-ethynyl-5α-androstane-3α,17β-diol.

FIG. 10 provides differential scanning calorimetry and thermogravimetricanalysis thermograms of a sample comprising Form I of17α-ethynyl-5α-androstane-3α,17β-diol.

FIG. 11 provides a Raman spectrum for a sample comprising Form I17α-ethynyl-5α-androstane-3α,17β-diol.

FIG. 12 provides an expanded view in a region of the Raman spectroscopyspectrum of FIG. 11.

FIG. 13 provides a comparison of calculated and experimentally derivedX-Ray powder diffraction patterns for Form I17α-ethynyl-5α-androstane-3α,17β-diol.

FIG. 14 is an ORTEP representation of the unit cell for crystalline FormI 17α-ethynyl-5α-androstane-3α,17β-diol determined from single crystalX-ray crystallography.

FIG. 15 provides a low resolution X-ray powder diffraction pattern of asample comprising Form IV 17α-ethynyl-5α-androstane-3α,17β-diol.

FIG. 16 provides differential thermal analysis and thermogravimetricanalysis thermograms of a sample comprising Form IV17α-ethynyl-5α-androstane-3α,17β-diol.

FIG. 17 provides a microscope photograph of crystals of Form IV17α-ethynyl-5α-androstane-3α,17β-diol under 2× and 10× magnification

FIG. 18 provides a low resolution X-ray powder diffraction pattern of asample comprising Form V 17α-ethynyl-5α-androstane-3α,17β-diol.

FIG. 19 provides a low resolution X-ray powder diffraction pattern of asample comprising Form VI 17α-ethynyl-5α-androstane-3α,17β-diol.

FIG. 20 provides differential thermal analysis and thermogravimetricanalysis thermograms of a sample comprising Form VI17α-ethynyl-5α-androstane-3α,17β-diol.

FIG. 21 provides a low resolution X-ray powder diffraction pattern of asample comprising Form VII 17α-ethynyl-5α-androstane-3α,17β-diol.

FIG. 22 provides an low resolution X-ray powder diffraction pattern of asample comprising Form VIII 17α-ethynyl-5α-androstane-3α,17β-diol.

FIG. 23 provides a low resolution X-ray powder diffraction pattern of asample comprising amorphous 17α-ethynyl-5α-androstane-3α,17β-diolsubstantially free of crystalline 17α-ethynyl-5α-androstane-3α,17β-diol.

FIG. 24 provides a thermogravimetric analysis thermogram and a thermaldifferential analysis thermogram of a sample comprising amorphous17α-ethynyl-5α-androstane-3α,17β-diol substantially free of crystalline17α-ethynyl-5α-androstane-3α,17β-diol.

FIG. 25 presents effects of a formulation prepared from crystalline17α-ethynyl-5α-androstane-3α,17β-diol on tumor incidence in a prostatecancer tumor model showing evolution of tumor volume by treatment andday since the start of treatment.

FIG. 26 presents effects of a formulation prepared from crystalline17α-ethynyl-5α-androstane-3α,17β-diol on established tumors in aprostate cancer model by showing the evolution of tumor volume bytreatment and day since the start of treatment.

FIG. 27 presents inhibition of cell cycle and induction of apoptosis inprostate cancer cells undergoing proliferation after treatment with aformulation prepared from crystalline17α-ethynyl-5α-androstane-3α,17β-diol.

FIG. 28 presents effects of a formulation prepared from crystalline17α-ethynyl-5α-androstane-3α,17β-diol on distribution of time to newtumor following first day of dosing in a breast cancer model

FIG. 29 presents effects of a formulation prepared from crystalline17α-ethynyl-5α-androstane-3α,17β-diol on tumor burden by volume in abreast cancer model with and without co-administration of TAXOTERE™ andby taxotere administered as a single agent.

FIG. 30 presents effects of ARIMIDEX™, TAMOXIFEN™ or vehicleadministered as a single agent in the breast cancer model of FIG. 29 forcomparison with the formulation prepared from crystalline17α-ethynyl-5α-androstane-3α,17β-diol administered with or withoutTAXOTERE™.

DETAILED DESCRIPTION Definitions

As used herein or otherwise stated or implied by context, terms that aredefined herein have the meanings that are specified. The descriptions ofembodiments and examples that are described illustrate the invention andthey are not intended to limit it in any way. Unless otherwisecontraindicated or implied, e.g., by mutually elements or options, inthe descriptions or throughout this specification, the terms “a” and“an” mean one or more and the term “or” means and/or.

Unless specified otherwise explicitly or by context, percentage amountsare expressed as % by weight (w/w). Thus, a solid-dosage formulationcontaining at least about 2% Compound 1 is a solid-dosage formulation orsuspension containing at least about 2% w/w Compound 1. Solid Compound 1containing 0.1% water means 0.1% w/w water is associated with the solid.

“About” and “approximately,” when used in connection with a numericvalue or range of values which is provided to describe a particularsolid form, e.g., a specific temperature or temperature range, such as,for example, that describing a melting, dehydration, desolvation orglass transition; a mass change, such as, for example, a mass change asa function of temperature or humidity; a solvent or water content, interms of, for example, mass or a percentage; or a peak position, suchas, for example, in analysis by IR or Raman spectroscopy or XRPD;indicate that the value or range of values may deviate to an extentdeemed reasonable to one of ordinary skill in the art while stilldescribing the particular solid state form. Specifically, the terms“about” and “approximately,” when used in this context, indicate thatthe numeric value or range of values may vary by 20%, 10%, 9%, 8%, 7%,6%, 5%, 4%, 3%, 2%, 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%,0.1% or 0.01% of the recited value or range of values while stilldescribing the particular composition or solid state form.

“Solid State” as used herein refers to a physical state of a compound orcomposition comprising the compound, such as17α-ethynyl-5α-androstane-3α,17β-diol (i.e., Compound 1); wherein atleast about 2-10% of the mass of the compound that is present exists asa solid. Typically, the majority of the mass of Compound 1 will be insolid state form. More typically, between at least about 80-90% of themass of Compound 1 is in solid form. Solid state forms includecrystalline, disordered crystalline, polycrystalline, microcrystalline,nanocrystalline, partially crystalline, amorphous and semisolid forms ormixtures thereof, optionally with non-solid or non-crystallineCompound 1. Solid state forms of Compound 1 further include polymorphs,pseudopolymorphs, hydrates, solvates, dehydrated hydrates and desolvatedsolvates and mixtures thereof, optionally with non-solid ornon-crystalline Compound 1. Thus, solid state forms of Compound 1 willinclude a single polymorph form of Compound 1, a single pseudo-polymorphform of Compound 1, a mixture of two or more, typically two or three,polymorph or pseudo-polymorph forms of Compound 1 or a combination ofany one of these solid state forms, optionally with non-solid ornon-crystalline Compound 1, provided that at least about 2-10% of themass of Compound 1 is in solid form.

The term “crystalline” and related terms used herein, when used todescribe a substance, component or product, means that the substance,component or product is crystalline as determined by a suitable method,typically X-ray diffraction [See, e.g., Remington's PharmaceuticalSciences, 18^(th) ed., Mack Publishing, Easton Pa., p 173 (1990); TheUnited States Pharmacopeia, 23^(rd) ed., pp. 1843-1844 (1995)].

The term “crystalline forms” and related terms herein refers to thevarious crystalline modifications of a given substance, including, butnot limited to, polymorphs, solvates, hydrates, mixed solvates,co-crystals and other molecular complexes. A crystalline form may alsobe a mixture various crystalline modifications of a given substance suchas a combination of pseudopolymorph or polymorph forms, a combination ofone or more polymorph forms with one or more pseudopolymorph or acombination of such forms with amorphous or non-solid state forms of thesubstance. Typical combinations with be of two or more polymorph orpseudo polymorph forms, such a mixture of a polymorph form with apseudopolymorph form or a mixture of a polymorph or pseudopolymorph formwith amorphous material.

Crystalline forms of a substance can be obtained by a number of methods,as known in the art. Such methods include, but are not limited to, meltrecrystallization, melt cooling, solvent recrystallization,recrystallization in confined spaces such as, e.g., in nanopores orcapillaries, recrystallization on surfaces or templates such as, e.g.,on polymers, recrystallization in the presence of additives, such as,e.g., co-crystal counter-molecules, desolvation, dehydration, rapidevaporation, rapid cooling, slow cooling, vapor diffusion, sublimation,grinding and solvent-drop grinding.

“Polymorph” as used herein refers to a defined crystalline form of asubstance such as Compound 1. Polymorphs typically differ in theirphysical properties due to the order of the molecules in the lattice ofthe polymorph. In addition, the physical properties of a polymorph candiffer, e.g., stability or flow characteristics, due to the presence ofhydrates, solvates or other molecules incorporated into the lattice ofthe polymorph.

In terms of thermodynamics, there are two types of polymorphism. Forexample, when polymorphs have a monotropic relationship, a plot of thefree energy of the various polymorphs in this relationship againsttemperature does not cross before all polymorphs melt, i.e., anytransition from one polymorph to another will be irreversible.Polymorphs that have a monotropic relationship are sometimes referred toas monotropes. For polymorphs having enantiotropic relationship, a plotof the free energy of the various polymorphs in this relationshipagainst temperature shows a crossing point before the various meltingpoints, and thus it may be possible to convert reversibly between thetwo polymorphs on heating and cooling. Polymorphs that have anenantiotropic relationship are sometimes referred to as enantiotropes.

Polymorphs may exhibit one or more differences in physical orpharmaceutical properties including hygroscopicity, solubility,intrinsic dissolution rate, solid state reaction rates (i.e., chemicalstability of a pharmaceutical ingredient as the drug substance or drugproduct), crystalline stability (i.e. tendency to transition to a morethermodynamically stable crystalline form), surface free energy,interfacial tension, mechanical strength (e.g., hardness, brittleness,plastic deformation, docility, malleability, etc.), tensile strength,compactability (i.e., tableting) and processability (e.g., handling,flow, blending, etc.). Differences in physical and mechanical propertiesof polymorphic forms of a drug substance may also affect scale-up andtransfer from laboratory procedures though pilot plant and then to fullproduction. Changes in equipment, variations in heating and coolingrates and variations in stirring procedures may also affectcrystallization and thus influence the crystalline form that isobtained.

Polymorphs existing as hydrates, solvates or mixed solvates aregenerally referred to as pseudopolymorphs and represent different solidstate forms in view of an isostructural polymorph form that is anhydrousor not a solvate. Pseudopolymorphs that differ in solvate identity orstoichiometry are also considered different solid state forms in view ofeach other. For example, Compound 1 existing as a monohydrate is adifferent solid state form in view of its isostructural dihydrate.Additionally, a methanol-water solvate (i.e., a mixed solvate) ofCompound 1 is viewed as a different solid state form in view of itsisostructural hydrate or anhydrate. Solvates and hydrates generallydemonstrate different solubilities or different intrinsic dissolutionrates compared to its isostructural anhydrate or desolvate. For example,a solvate may exhibit a lower intrinsic dissolution rate in the solventthat comprises the solvate as compared to its isostructural desolvate ata given temperature. Thus, a hydrate may sometimes exhibit a lowerintrinsic dissolution rate in an aqueous solution as compared to itsisostructural anhydrate. Furthermore, stability profiles of hydrates andsolvates at various temperatures and/or at different vapor pressures ofwater (e.g., relative humidity) or organic solvents will sometimesdiffer from those of the isostructural anhydrate or desolvate. Suchdifferences may influence formulation, processing or stability of anactive pharmaceutical ingredient (e.g., Compound 1), either as the drugsubstance or drug product under various storage conditions.

Thus, different crystalline or polymorphic forms may have differentphysical properties such as, for example, melting temperatures, heats offusion, solubilities, and/or vibrational spectra as a result of thearrangement or conformation of the molecules in the crystal lattice(see, e.g., Byrn, S. R., Pfeiffer, R. R., and Stowell, J. G. (1999)Solid-State Chemistry of Drugs, 2^(nd) ed., SSCI, Inc.: West Lafayette,Ind.). The differences in physical properties exhibited by polymorphsand pseudopolymorphs may affect pharmaceutical parameters such asstorage stability, compressibility and density (important in formulationand product manufacturing), and intrinsic dissolution rate, which can bean important factor in bioavailability. Differences in stability mayresult from changes in chemical reactivity (e.g., differentialoxidation, such that a dosage form discolors more rapidly when comprisedof one polymorph or pseudopolymorph than when comprised of anotherpolymorphic form) or mechanical changes (e.g., tablets crumble onstorage as a kinetically favored polymorph converts to thermodynamicallymore stable polymorph) or both (e.g., tablets of one polymorph are moresusceptible to breakdown at high humidity). As a result ofsolubility/dissolution differences, in the extreme case, somepolymorphic transitions may result in lack of potency or, at the otherextreme, toxicity. In addition, the physical properties of the crystalmay be important in processing, for example, one polymorph might be morelikely to form solvates or hydrates that may be difficult to filter andwash free of impurities (i.e., particle shape and size distributionmight be different between polymorphs).

Typically, crystalline forms are readily distinguished from each otherby one or more physical or analytical properties such as rate ofdissolution, Infrared and Raman spectroscopy, X-ray diffractiontechniques such as single crystal and powder diffraction techniques,solid state-NMR (SS-NMR), thermal techniques such as melting point,differential thermal analysis (DTA), differential scanning calorimetry(DSC), thermal gravimetric analysis (TGA) and other methods as disclosedelsewhere in the specification. Additional methods to characterize ordistinguish a pseudopolymorph from another isostructural polymorph,pseudopolymorph, desolvate or anhydrate include elemental analysis,Karl-Fisher titration, dynamic vapor sorption analysis,thermogravimetric-infrared spectroscopic analysis (TG-IR), residualsolvent gas chromatography, ¹H-NMR and other methods as disclosedelsewhere in the specification.

The term “isostructural crystalline form,” as used herein, refers to acrystal form of a substance that has a common structural similarity withanother crystalline form, including approximately similar interplanarspacing in the crystal lattice. [A more detailed account of crystallattices can be found in Chapters 2 and 3 of Stout and Jensen, X-RayStructure Determination: A Practical Guide, MacMillan Co., New York(1968)]. Thus, isostructural crystalline forms will have similarmolecular packing motifs, but differing unit cell parameters (a symmetrytranslation). Due to their common structural similarity, isostructuralcrystalline forms typically have similar, but not necessarily identical,X-ray powder diffraction patterns. An isostructural crystalline form maybe based upon a substance that is a neutral molecule or a molecularcomplex. The isostructural crystalline form may be a solvate, includinga hydrate, or a desolvated solvate crystalline form of the substance.Isostructural forms that are solvates of a polymorph are sometimesreferred to as pseudopolymorphic to the unsolvated polymorph. A solvatedcrystalline form typically contains one or more solvents, includingwater, in the crystal lattice, that may be the solvent or solvents ofcrystallization used in preparing the crystalline form.

“Amorphous”, as used herein, refers to a solid state form of a compound(e.g., Compound 1) wherein the solid state form has no long-rangeperiodic atomic structure as determined by X-ray powder diffraction(XRPD). The XRPD pattern of amorphous material will appear as a halowith no distinctive peaks. Amorphous material for some compounds can beobtained by a number of methods known in the art. Such methods include,but are not limited to, heating, melt cooling, rapid melt cooling,solvent evaporation, rapid solvent evaporation, desolvation,sublimation, grinding, cryo-grinding and freeze drying.

“Formulation” or “pharmaceutically acceptable formulation” as usedherein refers to a composition comprising an active pharmaceuticalingredient, such as 17α-ethynyl-5α-androstane-3α,17β-diol (i.e.,Compound 1), present in a solid state form in addition to one or morepharmaceutically acceptable excipients or refers to a compositionprepared from a solid state form of the active pharmaceuticalingredient, wherein the composition is suitable for administration to ahuman. The formulation may be comprised of, or be prepared from, one,two or more crystalline forms of the active pharmaceutical ingredient(e.g. a single polymorph or pseudopolymorph form of Compound 1, amixture of two polymorph forms of Compound 1, a mixture of a polymorphform of Compound 1 and a pseudopolymorph form of Compound 1 or a mixtureof a polymorph or pseudopolymorph form of Compound 1 and amorphousCompound 1. Typically, the formulations will be comprised of, orprepared from, a single crystalline form of Compound 1 (e.g., Form I,Form III, Form IV, Form V, Form VI, Form VII, Form VIII), amorphousCompound 1 or, less preferably, a mixture of a single polymorph orpseudopolymorph form and amorphous Compound 1.

“Solid state formulation” or “solid formulation” as used herein refersto a formulation comprising a solid state form of Compound 1 and one ormore pharmaceutically acceptable excipients wherein the majority of themass amount of the solid state form remains in that solid state form forat least about 6 months at ambient temperature, usually for at leastabout 12 months or 24 months at ambient temperature, when admixed withthe excipients in proportions required for the solid state formulation.Dosage units containing a solid state formulation include tablets,capsules, caplets, gelcaps, ampoules, suspensions and other dosage unitstypically associated with parenteral or enteral (oral) administration ofan active pharmaceutical ingredient in solid state form to a subject inneed thereof.

“Liquid formulation” as used herein refers to a formulation wherein oneor more solid state forms of Compound 1 has been admixed or contactedwith one or more excipients, wherein at least one of the excipients isin liquid or non-solid state form (i.e. a non-solid excipient), inproportions required for the liquid formulation, such that a majority ofthe mass amount of Compound 1 is dissolved into the non-solid excipient.Dosage units containing a liquid formulation include syrups, gels,ointments and other dosage units typically associated with parenteral orenteral administration of an active pharmaceutical ingredient to asubject in need thereof in non-solid state form.

“Substantially free” as used herein refers to a compound such asCompound 1 wherein more than about 60% by weight of the compound ispresent as the given solid state form. For example, the term“crystalline Compound 1 substantially free of amorphous material” refersto a solid-state form of Compound 1 wherein more than about 60% ofCompound 1 is crystalline Compound 1. Such compositions typicallycontain at least about 80%, preferably at least about 90%, ofcrystalline Compound 1 with the remaining present as amorphous ornon-crystalline Compound 1. Formulations described herein will typicallycontain about 94-99% of a single crystalline or amorphous form ofCompound 1, with about 97%, about 98% or about 99% preferred. In anotherexample, the term “amorphous Compound 1 substantially free ofcrystalline forms” refers to a solid-state form of Compound 1 whereinmore than about 60% of Compound 1 is amorphous. Such compositionstypically contain at least about 80%, usually at least about 90%,preferably at least about 95%, of amorphous Compound 1, with theremaining present as crystalline Compound 1. In yet another example, theterm “Form III substantially free of other crystalline forms” refers toa solid-state composition wherein more than about 60% of Compound 1exists in crystalline form as Form III. Such compositions typicallycontain at least about 80%, preferably at least about 90% and morepreferably at least about 97% of Compound 1 as Form III, with theremaining Compound 1 present as other crystalline or amorphous forms ora combination thereof.

“Substantially pure” as used herein refers to a solid state form ofCompound 1 that contain less than about 3% or less than about 2% byweight total impurities, or more preferably less than about 1% by weightwater, and/or less than about 0.5% by weight impurities such asdecomposition or synthesis by-products or residual organic solvent thatis not part of a solvate of a solid state form of Compound 1 (e.g. notpart of a pseudopolymorph) or other

“Substantially identical” as used herein refers to measured physicalcharacteristics that are comparable in value or data traces that arecomparable in peak position and amplitude or intensity with variationstypically associated with sample positioning or handling or the identityof the instrument employed to acquire the traces or physicalcharacteristics or due to other variations or fluctuations normallyencountered within or between laboratory environments or analyticalinstrumentation.

“Essentially free” as used herein refers to a component so identified asnot being present in an amount that is detectable under typicalconditions used for its detection or would adversely affect the desiredproperties of a composition or formulation in which the component may befound. For example, “essentially free of liquid” means a composition orformulation in solid form that does not contain water or solvent, inliquid form, in an amount that would adversely affect the pharmaceuticalacceptability of the formulation or composition for use in a soliddosage form to be administered to a subject in need thereof. Asuspension is considered a solid formulation and for such formulationsliquid excipient(s) comprising the suspension formulation are notincluded within this definition. “Polymorph Form III essentially free ofamorphous Compound 1” refers to a specific crystalline form of Compound1 in which amorphous Compound 1 is not detected by XRPD analysis.Typically, the detection limit for amorphous material within acrystalline form is about 10%.

“Hydrate” as used here refers to solid state form of a compound soidentified that contains water molecules as an integral part of thesolid state form and does not refer to water that is non-specificallybound to the bulk compound. Hydrates of Compound 1 in a crystalline formcan be isolated site hydrates or channel hydrates. In the crystalstructure of an isolated site hydrate the water molecules are isolatedfrom direct contact with other water molecules by the Compound 1molecules, whereas in channel hydrates the water molecules are locatednext to each other along one direction in the lattice. Hydrates cancontain stoichiometric or nonstoichiometric amounts of water moleculesper Compound 1 molecule. An expanded channel hydrate can take up waterinto the channels when exposed to high humidity and release water whenexposed to relatively low humidity. The crystal lattice of such hydratescan expand or contract as hydrate formation or dehydration proceeds,changing the dimensions of the unit cell. Typically, water will bepresent in a stoichiometric hydrate in the ratio of 0.25, 0.5, 1.0, 1.5or 2.0 relative to Compound 1. Hydrates are usually more stable thantheir anhydrous counterparts at conditions below its dehydrationtemperature. Isolated site hydrates usually dehydrate at relativelyhigher temperatures than channel hydrates. The dehydration process ofisolated site hydrates is sometimes destructive for the crystalstructure since it sometimes requires rearrangement of the molecules inthe unit cell in order to allow water molecules to escape the lattice.

“Solvate” as used here refers to solid state form of a compound soidentified that contains solvent molecules that is combined in adefinite ratio to the molecules of the compound and is an integral partof the solid state form and does not refer to solvent that isnon-specifically bound to bulk compound. When the solvent molecule iswater such solvates are referred to as hydrates. Typically, solvent willbe present in a solvate in the ratio of 0.25, 0.5, 1.0, 1.5 or 2.0relative to Compound 1.

“Hyperproliferation condition” or “cancer” as used here refers to acondition that is characterized by an abnormally high rate or apersistent state of cell division that is uncoordinated with that of thesurrounding normal tissues, and persists after, e.g., cessation of thestimulus that may have initially evoked the change in cell division.This uncontrolled and progressive state of cell proliferation may resultin a tumor that is benign, potentially malignant (premalignant) orfrankly malignant. Hyperproliferation conditions include thosecharacterized as a hyperplasia, dysplasia, adenoma, sarcoma, blastoma,carcinoma, lymphoma, leukemia or papilloma or other conditions describedherein.

“Hormone associated cancer, precancer or benign hyperplasia” or “hormonesensitive cancer, precancer or benign hyperplasia” as used herein refersto a hyperproliferation condition that responds negatively or positivelyin a therapeutic sense, to hormone manipulation or is a condition whosegenesis, persistence, invasiveness, refractivity, severity in symptomsor responsiveness to chemotherapy are attributable or related, in partor in whole, to hormone levels. Hormone associated or hormone sensitivecancers include, prostate cancer, breast cancer, ovarian cancer,cervical cancer, uterine cancer, endometrial carcinoma, adenocarcinoma,malignant melanoma or other conditions as described herein. Someadditional hormone associated or related cancers are described inMiller, A. B. Cancer Res. 38: 3985-3990 (1978).

Precancers are usually defined as lesions that exhibit histologicalchanges which are associated with an increased risk of cancerdevelopment and sometimes have some, but not all, of the molecular andphenotypic properties that characterize the cancer. Hormone associatedor hormone sensitive precancers include, prostatic intraepithelialneoplasia (PIN), particularly high-grade PIN (HGPIN), atypical smallacinar proliferation (ASAP), cervical dysplasia and ductal carcinoma insitu.

Hyperplasias generally refers to the proliferation of cells within anorgan or tissue beyond that which is ordinarily seen that may result inthe gross enlargement of an organ or in the formation of a benign tumoror growth. Hormone associated or hormone sensitive hyperplasias include,endometrial hyperplasia (endometriosis), benign prostatic hyperplasiaand ductal hyperplasia.

An “excipient”, “carrier”, “pharmaceutically acceptable carrier” orsimilar terms mean one or more component(s) or ingredient(s) that isacceptable in the sense of being compatible with the other ingredientsin compositions or formulations comprising Compound 1 as the activepharmaceutical ingredient that is in solid state form when admixed withthe excipients. These excipients usually are not overly deleterious to asubject to whom the composition formulation is to be administered. Asused here, “excipients” include liquids, such as water for injection,benzyl benzoate, cottonseed oil, N,N-dimethylacetamide, an alcohol suchas methanol, ethanol, glycerol, peanut oil, a polyethylene glycol(“PEG”), vitamin E, poppy seed oil, propylene glycol, safflower oil,sesame oil, soybean oil and vegetable oil. Excipients also includedissolution aids typically used for active pharmaceutical ingredientsthat are sparingly soluble or insoluble in water. Dissolution aidsinclude a cyclodextrin or a cyclodextrin derivative such asβ-cyclodextrin, 2-hydroxypropyl-β-cyclodextrin and CAPTISOL™ (sulfobutylether-β-cyclodextrin) and a PEG or PEG derivative such as CHREMOPHOR™ (apolyethoxylated castor oil). Any solid excipient may be a fine powder orgranulated. Excipients, as used herein may optionally exclude one ormore excipient, e.g., chloroform, dioxane, vegetable oil, DMSO, otherexcipients or any combination of these. Excipients include one or morecomponents typically used in the pharmaceutical formulation arts, e.g.,one, two or more of fillers, binders, disintegrants, dispersants,preservatives, glidants, surfactants and lubricants. Exemplaryexcipients include povidone, crospovidone, corn starch, carboxymethylcellulose, hydroxypropyl methylcellulose, microcrystalline cellulose,gum arabic, polysorbate 80, butylparaben, propylparaben, methylparaben,BHA, EDTA, sodium lauryl sulfate, sodium chloride, potassium chloride,titanium dioxide, magnesium stearate, castor oil, olive oil, vegetableoil, buffering agents such as sodium hydroxide, monobasic sodiumphosphate, dibasic sodium phosphate, potassium hydroxide, monobasicpotassium phosphate, dibasic potassium phosphate, tribasic potassiumphosphate, potassium carbonate, potassium bicarbonate, ammoniumhydroxide, ammonium chloride, saccharides such as mannitol, glucose,fructose, sucrose or lactose any of which may be compressible or any ofwhich may be spray dried.

A “subject” means a human or an animal. Usually the animal is a mammalor vertebrate such as a non-human primate dog or rodent. Subsets ofsubjects include subjects of a given species or group of species ofvarying ages, e.g., young humans, e.g., about 1 week of age to about 9years of age, adolescent humans, e.g., about 10-19 years of age, adulthumans, e.g., about 20-100 years of age, and mature adult or elderlyhumans, e.g., at least about 55 years of age, at least about 60 years ofage, at least about 65 years of age or a range of ages such as about60-100 years of age. Thus, as used herein, prevention or treatment of adisease, condition or symptom may include or exclude any subset ofsubjects that are grouped by age.

The terms “effective amount”, “effective dose” or the like generallymeans an amount of a solid state form of Compound 1 or an amount ofCompound 1 in a formulation comprised of or prepared from a solid stateform of Compound 1 that is sufficient to elicit a desired response,e.g., detectable restoration of normal physiological condition in asubject to which it is administered such as a decrease or stabilizationin tumor burden, detectable decrease of a cancer or hyperproliferationbiomarker, which may be a cell-surface biomarker or a circulatingbiomarker, a slowing in the rate of increase of the cancer orhyperproliferation biomarker or to detectable modulation or ameliorationof a cellular parameter or a clinical condition or symptom. An effectiveamount may be a single dose or two or more subdoses of Compound 1 in aformulation comprised of or prepared from a solid state form of Compound1 administered in one day, or it may be administered as multiple dosesover a period of time, e.g., over 2 days to about 1, 2, 3, 4 or 5 years.The effective amount may also be administered in multiple treatmentcycles as typically done in administration of cytotoxic agents for thetreatment of cancer. The treatment cycles may be separated by one ormore days or weeks, typically 1-4 weeks or may be separated by a longerperiod of time if remission of the hyperproliferation condition isachieved whereupon treatment is reinstituted upon recurrence of thecondition. Treatment cycles include daily administration of Compound 1for 4 weeks or 12 weeks.

“Prevent” or “prevention” of a condition or symptom as used here meansthat the onset of the condition or symptom can in some subjects bedelayed for at least some period of time in at least some treatedsubjects. “Prevent” or “prevention” can also be viewed as a delay indetectable dissemination of the hyperproliferation condition as measuredby delayed appearance of new lesions or metastasis. Such effects can beapparent in a significant minority of subjects (e.g., at least about 20%or more typically at least about 40%) or in a majority of subjects,which are observed in many clinical treatment situations, e.g., cancertreatments where a treatment can cause a disease to go into remissionand the remission can be permanent or exist for some period of time,e.g. about 1-3 months, about 4-6 months, about a year or about two tofive years. The treatments described herein can generate similareffects, which are referred to as preventing or prevention of thecondition or the symptom. Thus, “prevent” or “prevention” is notrestricted to keeping the occurrence of an event from ever happening orto preclude the possibility of the event from happening in all or amajority of all subjects, although such outcomes may occur.

“Subject to developing” as used herein refers to the likelihood of asubject, based upon risk factors predicated on pre-existing healthstatus, family history, behavior, genetic marker(s) or biochemicalmarker(s) that have been derived from a population of subjects to whichthe subject belongs, to suffer from a condition so identified. Thus, asubject, such as a human, who is subject to developing ahyperproliferation condition refers to a human subject with a generallyrecognized statistically greater likelihood of developing thehyperproliferation condition as a result of the human subject possessingone or more of the known risk factors for the hyperproliferationcondition. Individuals diagnosed with a precancer, e.g., cervicaldysplasia or prostatic intraepithelial neoplasia are considered to besubject to developing cervical cancer or prostate cancer.

A “surface-active agent” (surfactant) means a substance, which, at lowconcentrations, interacts between the surfaces of a solid and fluid inwhich the solid is insoluble or sparingly soluble. The fluid may be aliquid excipient present in a suspension formulation comprising a solidstate form of an active pharmaceutical ingredient, such as a crystallineForm I or Form III of Compound 1, the liquid excipient and the surfaceactive agent that acts to improve suspendability. Alternatively, thesurface active agent may be present in an oral solid dosage formcomprising a polymorph of Compound 1 (e.g., crystalline Form I or FormIII), the amorphous form of Compound 1 or a mixture thereof and thesurface active agent, which acts to improve dissolution rate of theactive pharmaceutical ingredient in the gastric fluid. Surface-activeagents are amphipathic in structure having both polar (hydrophilic) andnon-polar (hydrophobic) regions in the same molecule. Examples ofsurface active agents used in the formulation arts are given inCorrigan, O. I.; Healy, A. M. “Surfactants in Pharmaceutical Productsand Systems” in Encyclopedia of Pharmaceutical Technology 2^(nd) ed.Taylor and Francis, 2006, pp 3583-3596.

A “suspension” as used here unless specified or implied by context meansa solid state form of Compound 1 suspended, usually as a finely divided(e.g., micronized) solid, in a liquid carrier (vehicle) at a time priorto administration of the suspension. The suspension may be either readyto use or a dry powder reconstituted as a suspension dosage form justprior to use. Suspensions are used when Compound 1 is insoluble orpoorly soluble in a desired diluent or vehicle. Suspensions typicallyinclude a suspending or flocculating agent, a wetting agent, if thesuspending or flocculating agent that is present does not already servethis purpose, a buffering agent and a preservative. In a colloidalsuspension, the Compound 1 particles are typically less than about 1 μmin size. In a coarse suspension, they are larger than about 1 μm. Thepractical upper limit for individual suspendable Compound 1 particles incoarse suspensions is about 50 μm to 75 μm although some proportion ofparticles up to 200 μm may be suitable dependent upon the syringeabilityof the suspension. Design considerations for developing a suspension fororal or parenteral administration are given in Akers, et al. J.Parenteral Sci. Tech. 1987 41:88-96; Nash, RA “Suspensions” inEncyclopedia of Pharmaceutical Technology 2^(nd) ed. Taylor and Francis,2006, pp 3597-3610 (which is hereby incorporated herein by referencewith specificity into the present application).

Characterization and Identification Methods for Solid State Forms

Morphology—

Crystal morphology refers to the symmetry in a crystal as exhibited byits crystal faces due to the ordered internal arrangement of atoms inthe crystal structure. Crystal morphology of a particular crystallineform is typically described by the crystalline form's crystal system,namely, triclinic, monoclinic, orthorhombic, tetragonal, hexagonal orisometric. Crystal morphology may be determined by observation, forexample by microscopic evaluation under at least about 2×, 10× or 100×magnification using normal or polarized light. Crystals withcrystallographically distinct axes will interact with light in a mannerthat is dependent upon the orientation of the crystalline lattice withrespect to the incident light angle and are referred to as anisotropiccrystals. Thus, when light enters a non-equivalent axis, it is refractedinto two rays each polarized with the vibration directions oriented atright angles to one another, and traveling at different velocities. Thisphenomenon is termed birefringence and is exhibited to a greater orlesser degree in all anisotropic crystals. When polarized light isvibrating in a plane of the bifringent crystal that is parallel to thedirection of the polarizer there will be no contribution from lightpassing through the analyzer (because the single direction of lightvibration is parallel to the polarizer) resulting in the crystal beingvery dark and almost invisible (i.e., extinction). Thus, a bifringentcrystal will exhibit extinction when rotated under polarized light.Since many organic compounds in crystalline form are bifringent theircrystals will exhibit extinction provided that are well formed (i.e. arenot extensively fragmented or otherwise irregular in shape or containsignificant number of crystal defects). Therefore, a solid state form ofCompound 1 that does not exhibit extinction under examination withcross-polarized light does not necessarily mean that the solid stateform is not crystalline.

Crystal morphology can also be determined experimentally from singlecrystal X-ray data or computationally form X-ray powder diffraction databy methods disclosed herein.

X-Ray Powder Diffraction—

X-Ray powder diffraction (XRPD) is typically used to characterize oridentify crystalline compounds (see, e.g., U.S. Pharmacopoeia, volume23, 1995, method 941, p 1843-1845, U.S.P. Pharmacopeia Convention, Inc.,Rockville, Md.; Stout et al, X-Ray Structure Determination; A PracticalGuide, MacMillan Co., New York, N.Y. 1968). When an X-ray beam interactswith a crystalline form a diffraction pattern is typically producedcharacterized by sequences of intensity maximums at positions thatdepend on lattice features of the crystalline form. Thus, the positionsand the relative intensity of the XRPD lines are indicative of aparticular crystalline form that provide a “fingerprint” that is oftenspecific for a given crystalline form, although weak or very weakdiffraction peaks may not always appear in replicate diffractionpatterns obtained from successive batches of crystals. This isparticularly the case if other crystalline forms are present in thesample in appreciable amounts, e.g., when a polymorph or pseudopolymorphform has become partially hydrated, dehydrated, desolvated or heated togive a significant amount of another polymorph or pseudopolymorph form.

Furthermore, the relative intensities of bands, particularly at lowangle X-ray incidence values (low 2θ), may vary due to preferredorientation effects arising from differences in, e.g., crystal habit,particle size and other conditions of measurement. Thus, the relativeintensities of the diffraction peaks may not always be conclusivelydiagnostic of the crystal form in question. Instead, one typically looksto the relative positioning of the peaks coupled with their amplitude inorder to determine that a crystalline form of Compound 1 is one of theforms described herein. Broad XRPD peaks, which may consist of two ormore individual peaks located closely together, may be produced byamorphous components, disordered crystalline forms or parasitic scatterfrom the main beam. Broad peaks for different samples of the same solidstate form are generally located within about 0.3-1 degree 2θ. Sharpisolated XRPD peaks for different samples of the same solid state formare usually found for normal resolution data within about 0.1 2θ degreesor occasionally within about 0.2 2θ degrees on successive XRPD analyses,if they are conducted within the same lab under reproducibleenvironmental conditions following the same protocol. Thus, when a sharpisolated XRPD peak at a given position is identified as being locatedat, e.g., about 13.5 or 13.45 this means that the peak is at 13.5±0.1 or13.45±0.10. When a broad XRPD peak at a given position is identified asbeing located at about a given degree 2θ value, this means that the peakis at that degree 2θ value±0.3.

Under reproducible intra-lab conditions using the same instrument andprotocol to obtain the XRPD patterns, the differences in XRPD peaklocations and intensities obtained from successive XRPD analyses ondifferent samples of the same solid state form having the same degree ofcrystallinity are due primarily to differences in sample preparation orthe purity of the sample.

It is usually not necessary to rely on all bands that one observes in apurified polymorph or pseudopolymorph sample disclosed herein, sinceeven a single band may be diagnostic of a given polymorph orpseudopolymorph form of Compound 1. Rather, identification willtypically focus on band position and general pattern, particularly onthe selection of bands unique to the various polymorph andpseudopolymorph forms. Typically, an individual polymorph orpseudopolymorph form of Compound 1 is characterized by reference to the2, 3 or 4 most intense or the 2, 3 or 4 most reproducible peaks XRPDpeaks and optionally by reference to one or two other physical oranalytical properties such as melting point, one or more thermaltransitions observed in differential thermal analysis (DTA) and/ordifferential scanning calorimetry (DSC), one or more absorption peaksobserved in infrared or Raman spectroscopy and/or intrinsic dissolutionrate (DR) data in an aqueous or other solvent system. Standardizedmethods for obtaining XRPD, DTA, DSC, DR, etc. data have been describedfor example in U.S. Pharmacopoeia, volume 23, 1995, United StatesPharmacopeial Convention, Inc., Rockville, Md., pp 2292-2296 and2359-2765 (incorporated herein by reference).

One method to identify a known polymorph or pseudopolymorph form withina suspected solid state sample, such as a solid state formulationcomprising the known polymorph or pseudopolymorph form, involvesobtaining one or more XRPD patterns from sample(s) containing the knownpolymorph or pseudopolymorph form, which are then compared with the XRPDpatterns of the suspected solid state sample using, for example, aheuristic clustering analysis method as described for example in US Pat.Appl. No. 2004/0103130 (incorporated herein by reference particularly atparagraphs 0067-0078 and paragraphs 0086-0115 inclusive). Heuristicclustering analysis may also be used for quantitative analysis ofsamples containing either mixed crystalline phases (e.g., mixture of twoor more polymorph forms) or mixed crystalline and disordered phases(e.g. mixture of a polymorph and amorphous forms) as described forexample in US Pat. Appl. No. 2004/0103130 (incorporated herein byreference, particularly at paragraphs 0116-0130 inclusive).

Comparisons of atomic pairwise distribution functions (PDFs) derivedfrom XRPD patterns may also be used to identify a known polymorph orpseudopolymorph in a suspected solid state sample, such as a solid stateformulation comprising the known polymorph or pseudopolymorph form. Bydefinition, the PDF is the sine Fourier transform of the experimentallydetermined reduced structure factor obtained from a measured XRPDpattern and is obtained, for example, according to the procedure givenin Peterson, et al. “Improved measures of quality for the atomicpairwise distribution function” J. Acta Cryst. Vol. 36, pp. 53-64(2003). The PDF is an atomic density correlation function that describesthe solid state form by providing interatomic distances as given by thePDF peak positions and the number of atoms having a specific interatomicdistance as given by peak intensity. Thus, if two crystalline forms areof the same molecule with the same molecular packing, their PDFs will beessentially the same. To determine if two PDFs derived from, forexample, a known polymorph form or pseudopolymorph form and a solidstate formulation suspected of containing these crystalline forms areessentially identical, the PDFs are compared by, for example, the methoddescribed in US Pat. Appl. No. 2007/0243620 (incorporated herein byreference).

If high resolution XRPD pattern(s) of an essentially pure polymorph orpseudopolymorph may be obtained, then unit cell parameters (as describedin the section on single crystal X-ray analysis) may be determined forthe crystalline form by an indexing method as, for example, in US Pat.Appl. No. 2007/0270397 (incorporated herein by reference). For apseudopolymorph, if an isostructural crystalline form (i.e., a referencecrystalline form), such as an isostructural anhydrate, which may bederived from dehydration and/or desolvation of the pseudopolymorph, maybe obtained, then comparison of the unit cell volume of theisostructural crystalline form with the unit cell volume determined fromhigh resolution XRPD pattern(s) may allow determination of thestoichiometry of the pseudopolymorph (i.e., number of water or solventmolecules per molecule of Compound 1). In such applications, the unitcell parameters for the reference isostructural crystalline form may beobtained from single crystal X-ray analysis or derived from indexing ofhigh resolution XRPD data for this reference form.

An XRPD pattern may be described by “Prominent Peaks”, as is typicallydone for samples with only one XRPD pattern and limited other means toevaluate whether the sample provides a good approximation of the powderaverage. Prominent peaks are selected from observed peaks by identifyingpreferably non-overlapping, low-angle peaks, with strong intensity.

Single Crystal X-Ray Analysis—

Single X-ray crystallography identifies the smallest volume element,known as the unit cell that by repetition in three dimensions describesthe crystalline form. The dimensions of the unit cell is described bythree axes, a, b and c, and the angles between them α, β and γ. X-rayreflections from a series of planes are defined by the orientation andinterplanar spacings of these planes using three integers h, k and lcalled indices. A given set of planes with indices h, k, l cut thea-axis of the unit cell in h sections, the b-axis in k sections and thec-axis in l sections. A zero value for an indicia means the planes areparallel to the corresponding axis.

Single crystal X-ray parameters that characterize the crystalline formwill typically include the crystal system, space group, unit celldimensions, Z value (number of molecules per unit cell) and unit cellvolume. Typically, calculated density and ranges for indices h, k, l arealso used for characterization. Using the atomic coordinates, spacegroup and unit cell parameters determined from the single crystal dataone may simulate the XRPD pattern which may then be compared with theexperimentally determined XRPD pattern to confirm the correctness of thestructure solution for the unit cell.

Vibrational Spectroscopy—

Diagnostic techniques that one can optionally use to characterizecrystalline forms of Compound 1, such as a polymorph or pseudopolymorphform, include vibrational spectroscopy techniques such as IR and Raman,which measure the effect of incident energy on a solid state sample dueto the presence of particular chemical bonds within molecules of thesample that vibrate in response to the incident energy. Since themolecules in different polymorph or pseudopolymorph forms experiencedifferent intermolecular forces due to variations in conformational orenvironmental factors, perturbations of those vibrations occur thatleads to differences in spectra due to differences in frequency andintensity of some modes of vibration. Because polymorphs andpseudopolymorph form may possess different IR and Raman characteristicsfrom each other, IR and Raman spectrum provide complementary informationand either may provide a fingerprint for identification of a particularpolymorph. [see, Anderton, C. European Pharmaceutical Review, 9:68-74(2004)].

Raman is capable of determining polymorph or pseudopolymorph identityand/or quantification in a complex matrix, distinguishing betweenamorphous and crystalline forms or differentiating between multiplepolymorphic and pseudo polymorphic forms of Compound 1 [for example, seePratiwia, D., et al. “Quantitative analysis of polymorphic mixtures ofranitidine hydrochloride by Raman spectroscopy and principal componentsanalysis” Eur. J. Pharm. Biopharm. 54(3), 337-341 (2002)]. Foridentifying a polymorph or pseudopolymorph form in a solid formulationsuch as a tablet, powder samples of these pure crystalline forms ofCompound 1 and excipients are gently compacted and scanned with Ramanmicroscopy to build up a library of formulation component spectra. Apartial least squares (PLS) model and multivariate classification arethen used to analyze Raman mapping data obtained from sectioned tabletshaving low API content (about 0.5% w/w). Multivariate classificationallows polymorph assignments to be made on individual microscopic pixelsof Compound 1 identified in the data. By testing data from separate setsof tablets containing each specific crystalline form, specific formrecognition may be demonstrated at about 0.5% w/w. For tabletscontaining a mixture of crystalline forms, recognition of about 10%polymorphic or pseudopolymorphic impurity of Compound 1 (representing anabsolute detection limit of about 0.05% w/w), is possible.

For determining polymorph or pseudopolymorph identity or quantificationfor a crystalline form of Compound 1 within a complex matrix such as asolid formulation using the above vibrational spectroscopy methods, thetechnique of attenuated total reflectance (ATF) is sometimes used (foran example see Salari, H., et al. “Application of attenuated totalreflectance FTIR spectroscopy to the analysis of mixtures ofpharmaceutical polymorphs” International Journal of Pharmaceutics 163(1): 157-166 (1998)].

Another technique for identification or quantification of a crystallineform is Diffuse Reflectance Infrared Fourier Transform Spectroscopy(DRIFTS) (for an example see Tantishaiyakul, V., et al. “Use of DRIFTSand PLS for the Determination of Polymorphs of Piroxicam alone and incombination with pharmaceutical excipients: A Technical Note” AAPSPharmSciTech 9(1) 95-99 (2008)]. It is well known that particle size isa key variable in diffuse reflectance measurements, since largeparticles will result in scattering of the energy leading to the shiftof the spectrum baseline and the broadening of IR bands. To conductDRIFTS the sample containing solid state Compound 1 is prepared, bygrinding or passing it through a sieve, to obtain uniform particles withthe later preferred since the possibility of transformation of ametastable polymorph or pseudopolymorph form to another crystalline formis avoided.

In yet another technique, near-infrared (NIR) spectroscopy may also beused in identification or quantitative analysis of a crystalline form,such as a polymorphs or pseudo polymorph form (e.g. hydrate) of Compound1 in mixture of solid state forms or identification of a polymorph orpseudopolymorph form in a solid formulation such as a tablet containingthe polymorph or pseudopolymorph form of Compound 1.

Extensive overlap of IR or Raman bands from different crystalline formsof Compound 1 examined by the various vibration spectroscopy methodsdescribed herein may sometimes occur so that quantification requiresdeconvolution methods to extract information for each individualcomponent. Such deconvolution methods include partial least squaresregression, principle component analysis or other methodologies [forexamples, see Reich, G. “Near-infrared spectroscopy and imaging: Basicprinciples and pharmaceutical applications” Adv. Drug Deliv. Rev. 57:1109-43 (2005)].

Solid State Nuclear Magnetic Resonance (SS-NMR)—

Diagnostic techniques that one can optionally use to characterizepolymorphs of Compound 1 include solid state NMR techniques [forexamples see Tishmack, P. A., et al. “Solid-State Nuclear MagneticResonance Spectroscopy: Pharmaceutical Applications,” J. Pharm. Sci. 92(3): 441-474 (2003)]. These techniques offer the advantage of beingnondestructive and noninvasive. SS-NMR spectroscopy is sometimessuitable for testing drug formulations, such as those comprisingCompound 1, because the NMR resonances for most pharmaceuticalexcipients occur in a narrow range of the NMR spectrum. Thus, it istypically easy to distinguish excipients from Compound 1 NMR resonances.Spectral subtraction can even be used to eliminate interfering excipientpeaks that are present from the spectrum.

SS-NMR usefulness in characterizing a particular polymorph orpseudopolymorph form of Compound 1 is due to different numbers ofcrystallographically non-equivalent sites in the unit cells of thesecrystalline forms, and its sensitivity to changes in the local chemicalenvironment where slight changes in bond lengths, bond angles,interactions with neighboring molecules or, in the case of pseudopolymorphs, different hydration or solvation levels, can effect theSS-NMR signals relative to the solution spectrum.

For amorphous materials, SS-NMR spectroscopy may be used to examine thedegree of disorder because various processing techniques (e.g.,lyophilization, spray drying, melt-quench, cryomilling) can vary theoverall degree of disorder in the sample or result in a change inpolymorph form. For a less stable (i.e., metastable) polymorph orpseudopolymorph form of Compound 1, the relative instability of thatcrystalline form could be caused by the overall greater molecularmobility of Compound 1 in the solid state sample.

Sometimes, the same polymorphic or pseudopolymorphic form of a compoundobtained from different lots may exhibit different physiochemicalproperties such as stability and dissolution rate, which could be causedby the degree of crystallinity in the sample. The presence of defectsites or less crystalline domains in the solid state sample causing aloss of degree of crystallinity sometimes may not be observed in X-raypowder diffraction. This is often not the case with SS-NMR spectroscopy,since these sites or domains provide another avenue for the relaxationprocess of the spin states in the solid state sample.

SS-NMR may also be applied to analyzing solid formulations comprisingCompound 1 and thus may be useful for detecting different solid stateforms of Compound 1 in the presence of excipients. For detectingamorphous Compound 1 the detection limit for SS-NMR is sometimes about10-20%, depending on the relative location of the amorphous andcrystalline Form peaks in the spectrum, because amorphous peaksgenerally are very broad. This is about the same detection limit forXRPD.

SS-NMR spectroscopy is suitable for testing drug formulations becausethe NMR resonances for most pharmaceutical excipients occur in a narrowrange of the NMR spectrum. Thus, distinguishing NMR resonances ofCompound 1 from excipients in a formulation containing a particularpolymorph or pseudopolymorph form of Compound 1 is typically possible.If there are interfering excipient peaks in the spectrum, spectralsubtraction may also be used to eliminate or reduce this interference.

For identification of a particular crystalline form of Compound 1 withina formulation, SSNMR is sometimes superior to XRPD, since NMR peaksassociated with Compound 1 may be found that are not obscured by peaksfrom excipients of the formulation. This may not be the case in XRPDsince many diffraction lines may overlap, thus limiting the detection ofsmall amounts of a polymorph or pseudopolymorph form of Compound 1.Without interfering diffraction lines a detection limit of about 10%,and sometimes to about 5%, in XRPD for a Compound 1 polymorph orpseudopolymorph may be obtained, while SSNMR may attain detection limitsdown to about 0.5%. In addition, because NMR spectroscopy is inherentlya quantitative technique (i.e., signal intensity is relative to thenumber of nuclear sites at that specific resonance frequency), SS-NMRspectroscopy may allow one to determine the contribution of crystallineforms of Compound 1, or of crystalline and amorphous Compound 1, in amixture of such forms.

Thermal Analysis Procedures—

Diagnostic techniques that one can optionally use to characterizepolymorphs of Compound 1 include differential thermal analysis (DTA),differential scanning calorimetry (DSC), thermogravimetric analysis(TGA) and melting point measurements.

DTA and DSC measures thermal transition temperatures at which acrystalline form absorbs or releases heat when its crystal structurechanges or it melts. TGA is used to measure thermal stability and thefraction of volatile components of a sample by monitoring the weightchange as the sample is heated. If infrared spectroscopy is conducted onthe volatile components outgassed during TGA analysis of apseudopolymorph (TGA-IR), then the molecular composition of thepolymorph may be discerned. These techniques are thus useful forcharacterizing solid state forms existing as solvates and/or hydrates.

DTA involves heating a test sample of a solid state form of Compound 1and an inert reference under identical conditions while recording anytemperature difference between the sample and reference. Thisdifferential temperature is plotted against temperature, and changes inthe test sample that leads to absorption or liberation of heat can thusbe determined relative to the inert sample.

DSC measures the energy needed to establish a nearly zero temperaturedifference between a sample and an inert reference as they are subjectedto identical heating regimes. The energy required to do this is ameasure of the enthalpy or heat capacity changes in the sample relativeto the reference.

Thermal transition temperatures typically occur within about 2° C. onsuccessive analyses using a temperature scan rate of 10° C./min andoccur within about 1 degree depending on the temperature scan rate used(with slower scan rates such as 5° C./min or 1° C./min providing greaterprecision). When it is stated that a compound has a DTA transition at agiven value, it means that the DTA transition will be within ±2° C.Different crystalline forms including polymorph or pseudopolymorph formsmay be identified, at least in part, based on their different transitiontemperature profiles in their DTA thermographs.

Thermal analysis is usually conducted at a temperature scan rate of 10°C./min. Lower scan rates such as 5° C./min or 1° C./min may be used ifoverlap of temperature transitions is suspected. Thus, a suspectedtransition due to a change in polymorph form to a different, more stablepolymorph prior to complete melting of the sample may be discerned usinga slower scan rate. A transition during thermal analysis of akinetically formed polymorph to a thermodynamically more stablepolymorph prior to complete melting may be avoided using a faster scanrate that does not allow time for the transition to occur.

Data Acquisition for Characterization and Identification Methods

Data provided in the Figures, Tables and Examples were obtained usingthe following methods and instrumentation.

X-Ray Powder Diffraction—

XRPD patterns were obtained using a PANalytical X'Pert Prodiffractometer. An incident beam of Cu Kα radiation was produced usingan Optix long, fine-focus source. An elliptically graded multilayermirror was used to focus the Cu Kα X-rays of the source through thespecimen and onto the detector. Data were collected and analyzed usingX'Pert Pro Data Collector software (v. 2.2b). Prior to the analysis, asilicon specimen (NIST SRM 640c) was analyzed to verify the Si 111 peakposition. The specimen was sandwiched between 3 μm thick films, analyzedin transmission geometry, and rotated to optimize orientationstatistics. A beam-stop and a helium atmosphere were used to minimizethe background generated by air scattering. Soller slits were used forthe incident and diffracted beams to minimize axial divergence.Diffraction patterns were collected using a scanning position-sensitivedetector (X'Celerator) located 240 mm from the specimen.

XRPD patterns were collected using an Inel XRG-3000 diffractometerequipped with a curved position sensitive detector with a 2θ range of120°. An incident beam of Cu Kα radiation (40 kV, 30 mA) was used tocollect data in real time at a resolution of 0.03° 2θ. Prior to theanalysis, a silicon standard (NIST SRM 640c) was analyzed to verify theSi 111 peak position. Samples were prepared for analysis by packing theminto thin-walled glass capillaries. Each capillary was mounted onto agoniometer head and rotated during data acquisition. The monochromatorslit was set at 5 mm by 160 μm, and the samples were analyzed for 5minutes.

XRPD patterns were also obtained on a Shimadzu WRD-6000 X-ray powderdiffractometer with Cu Kα radiation. The solid state samples wereprepared for analysis by placement in an aluminum holder with a siliconinsert. The instrument was equipped with a long fine focus X-ray tube.The tube voltage and amperage were set to 40 kV and 40 mA, respectively.The divergence and scattering slits were set at 1° and the receivingslit was set at 0.15 mm. Prior to the analysis, a silicon standard (NISTSRM 640c) was analyzed to verify the Si 111 peak position. Diffractionradiation was detected by a sodium iodide scintillation detector. A θ-2θcontinuous scan at 3°/min (0.4 sec/0.02° step) from 2.5 to 40° 2θ wasused.

X-ray diffraction patterns presented herein are accompanied by labeledpeaks and tables with peak lists. Reported peak data, under mostcircumstances, is within the range of up to about 30° 2θ. As previouslydiscussed data will be instrument dependent and third party measurementson independently prepared samples on different instruments may lead tovariability that is greater than ±0.1° 2θ. In addition, for technicalreasons, different rounding algorithms were used to round each peak tothe nearest 0.1° or 0.01° 2θ, depending upon the instrument used tocollect the data and/or the inherent peak resolution.

The location of reported peaks along the x-axis (degree 2θ) in thefigures and the tables were automatically determined using PATTERNMATCH™2.4.0 software and rounded to one or two significant figures after thedecimal point based upon the above criteria. Peak position variabilityis given to within ±0.1° 2θ based upon recommendations outlined in theUSP discussion of variability in X-ray powder diffraction given inUnited States Pharmacopeia, USP 31, NF 26, Vol. 1, pg. 374. For d-spacelistings, the wavelength used to calculate d-spacings was 1.541874 Å, aweighted average of the Cu-K_(α1) and Cu-K_(α2) wavelengths [Phys. Rev.A56(6) 4554-4568 (1997)]. Variability associated with d-spacingestimates was calculated from the USP recommendation at each d-spacingand is provided in the respective data tables.

Differential Scanning Calorimetry (DSC)—

DSC was performed using a TA Instruments Q2000 differential scanningcalorimeter. Temperature calibration was performed using NIST traceableindium metal. The sample was placed into an aluminum DSC pan, and theweight was accurately recorded. The pan was covered with a lidperforated with a laser pinhole, and the lid was crimped. A weighed,crimped aluminum pan was placed on the reference side of the cell. Thesample cell was equilibrated at 25° C. and heated under a nitrogen purgeat a rate of 10° C./minute, up to a final temperature of 250° C.Reported temperatures are at the transition maxima.

Differential Thermal Analysis (DTA)—

DTA and TGA were performed simultaneously using a Seiko SSC 5200 TG/DTAinstrument. Temperature calibration was performed using NIST traceableindium metal. The sample was placed into an aluminum pan and looselycovered with a lid and the weight accurately recorded. The sample cellwas equilibrated at 25° C. and then heated under a nitrogen purge at arate of 10° C./minute, up to a final temperature of 250° C. Reportedtemperatures are at the transition maxima.

Thermogravimetric Analysis (TGA)—

TGA was performed using a TA Instruments Q5000 IR thermogravimetricanalyzer. Temperature calibration was performed using nickel andALUMEL™. Each sample was placed in an aluminum/or/platinum pan. The panwas hermetically sealed with a lid that was opened using a punchingmechanism just before being inserted into the TG furnace. The furnacewas heated under nitrogen at a rate of 10° C./minute to a finaltemperature of 350° C.

FT-Raman Spectroscopy—

Raman spectra were acquired on a Nexus 670 FT-Raman accessory moduleinterfaced to a Nexus 670 FT-IR spectrophotometer (Thermo Nicolet)equipped with an indium gallium arsenide (InGaAs) detector. Wavelengthverification was performed using sulfur and cyclohexane. Each sample wasprepared for analysis by placing the sample into a glass tube andpositioning the tube in a gold-coated tube holder. Approximately 0.5 Wof Nd:YVO₄ laser power (1064 nm excitation wavelength) was used toirradiate the sample. Each spectrum represents 256 co-added scanscollected at a spectral resolution of 4 cm⁻¹.

Karl Fischer Analysis—

Coulometric Karl Fischer (KF) analysis for water determination wasperformed using a Mettler Toledo DL39 KF titrator. A blank titration wascarried out prior to analysis. The sample was prepared under a drynitrogen atmosphere, and dissolved in approximately 1 mL dryHYDRANAL-COULOMAT AD™ in a pre-dried vial. The entire solution was addedto the KF coulometer through a septum and mixed for 10 seconds. Thesample was then titrated by means of a generator electrode, whichproduces iodine by electrochemical oxidation: 2 l⁻→l₂+2e⁻. Tworeplicates were obtained to ensure reproducibility.

Formulations—

Formulations comprising Compound 1 as the active pharmaceuticalingredient will have a significant percentage of Compound 1 in one ormore of its solid state forms, typically in one or two solid stateforms. Exemplary formulations contain at least about 60% or usually atleast about 90% of Compound 1 in one solid state form. Formulations willusually comprise one or more given solid state forms of Compound 1,substantially free of other solid state forms, and one or more,typically 1, 2, 3 or 4 excipients or carriers. Other formulations cancontain Compound 1 in one or more solid state forms, typically one ortwo. Other formulations are generally solids, precipitates, gels,suspensions and colloids that contain one or more solid state forms ofCompound 1, such as the amorphous form of Compound 1, crystalline Form Ior crystalline Form III of Compound 1 or a mixture thereof. Preferredoral unit dosages for human use will contain about 2 mg, 5 mg, 10 mg, 15mg, 20 mg or 40 mg of a solid state form of Compound 1 per unit dose.

While it is possible to administer Compound 1 in its solid state as apure compound to a subject, it is usually presented as a solidformulation essentially free of liquid or less frequently a solidsuspension. Formulations will typically be used to prepare unit dosages,e.g., tablets, capsules or lozenges for oral, buccal or sublingualadministration, that usually comprise about 0.1-500 mg, typically about0.5-100, or about 1-100 mg (e.g., about 0.1, about 0.25, about 0.5,about 1, about 5, about 10, about 20, about 100 mg) of a formulationcontaining a solid state form of Compound 1 such as amorphous Compound1, crystalline Form I or crystalline Form III of Compound 1.Alternatively, embodiments include a product for parenteral (e.g.,subcutaneous, subdermal, intravenous, intramuscular, intraperitoneal oraerosol) administration made by the process of contacting a solid stateform of Compound 1, such as amorphous Compound 1, crystalline Form I, orcrystalline Form III of Compound 1, with a liquid excipient, e.g., anyone, two, three or more of PEG 100, PEG 200, PEG 300, PEG 400, propyleneglycol, benzyl benzoate, benzyl alcohol or ethanol, and optionallysterilizing the solution and optionally dispensing the solution intovials or ampoules (typically amber glass), which may be single-use ormulti-use and optionally storing the formulation at reduced temperature(about 0-12° C., or about 2-10° C.). Such products for parenteraladministration typically comprise Compound 1 at a concentration of about0.1-100 mg/mL, usually at about 1-100 mg/mL or about 10-100 mg/mL, andoptionally containing one or more of a salt, buffer or bacteriostat orpreservative (e.g., NaCl, BHA, BHT or EDTA). Sometimes a surface activeagent is used to affect a suspension or is incorporated into an oralsolid dosage form to assist dissolution of the solid state form ofCompound 1 into the gastric tract. In general, formulations for oraladministration are preferred for human therapeutic applications.

Surface active agents used in a suspension or a solid form of Compound 1in a liquid excipient(s) include nonionic, cationic and anionicsurfactants. Examples of preferred surfactants include, but are notlimited to, sodium lauryl sulfate, sodium dodecyl sulfate, polysorbate40 and polysorbate 80.

In one embodiment, sodium lauryl sulfate is used as a surface activeagent in a unit dosage form, such as a tablet or a capsule, for oraladministration in treatment of a condition disclosed herein wherein theformulation comprises crystalline Form I or crystalline Form III ofCompound 1 essentially free of other solid state forms of Compound 1 andthe surface active agent, optionally comprising one or more additionalexcipients, typically 1, 2 or 3 additional excipients.

Examples of other excipients used in the preparation of formulationscomprising a solid state form of Compound 1 (e.g., crystalline Form I orcrystalline Form III), by way of illustration and not limitation, aregiven in Nema, S., et al. PDA J. Pharm. Sci. Tech. 1997, 51:166-171;Strickley, R. G. Pharm. Res. 2004, 21:201-230; Powell, M. F., et al. PDAJ. Pharm. Sci. Tech 1998, 52:238-311; Akers, M. J. in “Drug Delivery:Parenteral Route” Encyclopedia of Pharmaceutical Technology, InformaHealthcare, USA, 2007, pp 1266-1278.

Micronization—

To improve dissolution rate of a crystalline form of Compound 1 in aformulation comprising at least one crystalline form of Compound 1 andone or more pharmaceutically acceptable excipients in a solid dosageform or to affect suspendability in a suspension for oral or parenteraladministration comprising a crystalline form of Compound 1 and a liquidexcipient(s), the crystalline form may be milled to an mean volumeweighted particle size (Dv,50) or average diameter of about 0.01-200 μm,or about 0.1-100 μm or about 3-50 μm. Mean volume weighted particle size(Dv, 50) or average diameter for milled crystalline Compound 1 may thusbe relatively small, e.g., about 0.03-2.0 μm or about 0.1-1.0 μm, orsomewhat larger, e.g., about 3-100 μm, about 5-75 μm or about 10-30 μm.Milled crystalline Compound 1 is suitable for preparing solid andsuspension formulations intended for oral or parenteral administrationto a subject. Mean volume weighted particle size or average diameterinclude a range between about 0.01 and about 500 microns in 0.05 micronor in 0.1 micron, e.g., mean volume weighted particle size or averagediameter of about 0.05, about 0.1, about 0.5, about 1.0, about 1.5,about 2.0, about 2.5, about 3.0, about 3.5, about 4.0, about 4.5, about5.0, about 5.5, about 6, about 7, about 8, about 9, about 10, about 15,about 20, about 25, about 30, about 35, about 40, about 50, about 60,about 75, about 85, about 100, about 120, etc. microns). Preferably,mean volume weighted particle size (Dv,50) or average diameter are about5, about 10, about 15 or about 20 micron. The particle size (Dv, 90)typically is about 5 micron, about 10, about 15, about 20, about 25 orabout 30 micron. Preferred particle size has (Dv, 90) of ≦10 microns orabout micron (Dv, 90) of ≦6.8 microns.

As used herein, reference to a mean volume weighted particle size oraverage diameter means that the material, e.g., crystalline compound 1,an excipient(s) or a composition that comprises both, is ground, milled,sieved or otherwise treated so as to comprise the specified particlesize. It is to be understood that some particles may be larger orsmaller (i.e., will exist in a distribution of particle sizes), but thecomposition or the crystalline form of Compound 1 (e.g. crystalline FormI or crystalline Form III) will comprise a significant proportion of thematerial with the specified size or within an acceptable range of thespecified size. Micronization methods include milling by ball mills, pinmills, jet mills (e.g., fluid energy jet mills) and grinding, sievingand precipitation of a compound(s) from a solution, see, e.g., U.S. Pat.Nos. 4,919,341, 5,202,129, 5,271,944, 5,424,077 and 5,455,049, which arespecifically incorporated herein by reference herein). Particle size isdetermined by, e.g., transmission electron microscopy, scanning electronmicroscopy, light microscopy, X-ray diffractometry and light scatteringmethods or Coulter counter analysis (see, for example, “Characterizationof Bulk Solids” D. McGlinchey, Ed., Blackwell Publishing, 2005).

Thus, crystalline Compound 1 may comprise or consist essentially of apowder that contains one, two or more of these mean volume weightedparticle sizes or average diameter particle sizes and the powder may becontacted with a solid excipient(s), which can be mixed and optionallycompressed or formed into a desired shape. Alternatively, crystallineCompound 1 formed into a powder a described above is contacted with aliquid excipient(s) to prepare a liquid formulation or a liquidcomposition that is incorporated into a solid formulation or suspension.Suitable micronized formulations thus include aqueous or oilysuspensions of crystalline Compound 1.

Hyperproliferation Conditions—

Hyperproliferation conditions that can be treated with crystallineCompound 1, for example, an anhydrate crystalline form such as Form III,or amorphous Compound 1 include cancers, precancers and otherhyperproliferation conditions comprising carcinomas, sarcomas, adenomas,dysplasias, blastomas, papillomas, naevus, pre-malignant tumors, benigntumors or malignant tumors including solid tumors and disseminatedtumors such as one associated with or arising from prostate, lung,breast, ovary, skin, stomach, intestine, pancreas, neck, larynx,esophagus, throat, tongue, lip, oral cavity, oral mucosa, salivarygland, testes, liver, parotid, biliary tract, colon, rectum, cervix,uterus, vagina, pelvis, endometrium, kidney, bladder, central nervoussystem, glial cell, astrocyte, squamous cell, blood, bone marrow, muscleor thyroid cells or tissue.

One category of benign tumors encompasses functional tumors so namedbecause they have functional effects on the affected tissue. Forexample, functional tumors of endocrine tissue, referred to as adenomas,overproduce certain hormones.

Another category of benign tumors that can be treated with crystallineCompound 1, for example, an anhydrate crystalline form such as Form III,or amorphous Compound 1 includes papillomas, which refer to benignepithelial tumors growing exophytically (outwardly projecting) infinger-like fronds, and naevus. Papillomas include Larynx papilloma,Choroid plexus papilloma, skin papilloma, squamous cell papilloma andtransitional cell papilloma (also known as bladder papilloma).

Other benign tumors that can be treated with crystalline Compound 1, forexample, an anhydrate crystalline form such as Form III, or amorphousCompound 1 are cystadenoma (or “cystoma”), which is a type of cysticadenoma derived from glandular tissue where secretions are retained andaccumulate in cysts and include mucinous cystadenoma (produced byovarian epithelial cells), papillary cystadenoma (any tumor thatproduces patterns that are both papillary and cystic), serouscystadenoma and thecomas, which are benign ovarian neoplasms that aretypically estrogen-producing.

Non-malignant hyperproliferative conditions of the skin that can betreated with crystalline Compound 1, for example, an anhydratecrystalline form such as Form III, or amorphous Compound 1 includeseborrheic keratosis, toxic eczema, allergic eczema, atopic dermatitis,ichthyosis, and psoriasis.

Benign prostate hyperplasia (BPH), arteriovenous malformations,heterotrophic bone formation, hyperplasia of the breast, focalepithelial hyperplasia, sebaceous hyperplasia and congenital adrenalhyperplasia are examples of hyperproliferation conditions that arehyperplasias that can be treated with crystalline Compound 1, forexample, an anhydrate polymorphic form such as Form III, or amorphousCompound 1. Preferable treatments are for BPH.

Treatment options for BPH include crystalline Compound 1, for example,an anhydrate crystalline form such as Form III, or amorphous Compound 1include α₁-adrenergic receptor antagonists such as doxazosin, terazosin,alfuzosin, tamsulosin and 5α-reductase inhibitors such as finasterideand dutasteride.

Premalignant conditions or tumors that can be treated with crystallineCompound 1, for example, an anhydrate crystalline form such as Form III,or amorphous Compound 1 include colon polyps, actinic keratosis,squamous metaplasia, leukoplakia, erythroplakia, Barrett's esophagus,endometrial hyperplasia, cervix dysplasia, polycythemia rubra vera andcarcinoma in situ (CIS). Preferably treated are endometrial hyperplasiaand cervix displasia

Prostatic interstitial neoplasia (PIN) can be treated with crystallineCompound 1, for example, an anhydrate crystalline form such as Form III,or amorphous Compound. PIN can be classified as high grade, medium gradeand low grade.

Dysplasia (or heteroplasia) that can be treated with crystallineCompound 1, for example, an anhydrate crystalline form such as Form III,or amorphous Compound 1 refers to an abnormality in maturation of cellswithin a tissue and include myelodysplastic syndromes or dysplasia ofblood-forming cells.

Malignant tumors (cancers) have properties that allow invasion anddestruction of nearby tissue and spreading (metastasizing) to otherparts of the body. Cancers are classified by the type of cell thatresembles the tumor and, therefore, the tissue presumed to be the originof the tumor. One general category of cancer that can be treated withcrystalline Compound 1, for example, an anhydrate crystalline form suchas Form III, or amorphous Compound 1 encompasses the carcinomas, whichinclude common forms of breast, prostate, lung and colon cancer as wellas basal cell carcinoma, malignant melanoma, squamous cell carcinoma,which is a malignant tumor of squamous epithelium, and also occurs insites including the lips, mouth, esophagus, urinary bladder, prostate,lungs, vagina and cervix. Preferred treatments are for prostate andbreast carcinoma. Also preferred are treatments for lung and cervixcarcinoma.

Adenocarcinomas that can be treated with crystalline Compound 1, forexample, an anhydrate crystalline form such as Form III, or amorphousCompound 1 represent a category of carcinomas and are of glandularorigin (vide supra). To be classified as adenocarcinoma, the cells donot necessarily need to be part of a gland, as long as they havesecretory properties. Adenocarcinomas sometimes begin in cells lininginternal organs that have gland-like (secretory) properties and thus mayarise in numerous tissues including those of breast, colon, lung,prostate, pancreas, stomach, cervix and vagina.

Carcinomas that can be treated with crystalline Compound 1, for example,an anhydrate crystalline form such as Form III, or amorphous Compound 1include renal cell carcinoma, endometrial carcinoma and hepatocellularcarcinoma (HCC), also called hepatoma. Preferred treatments are for HCCand endometrial carcinoma.

Another category of malignant tumors that also can be treated withcrystalline Compound 1, for example, an anhydrate crystalline form suchas Form III, or amorphous Compound 1 encompasses the neuroendocrinetumors which include insulinoma and glucagonoma.

Another category of malignant tumors that can be treated withcrystalline Compound 1, for example, an anhydrate crystalline form suchas Form III, or amorphous Compound 1 are sarcomas, including lymphomas,leukemias, germ cell tumors and blastomas such as glioblastoma andmedulloblastoma, hepatoblastoma, nephroblastoma, neuroblastoma,osteoblastoma and retinoblastoma. Preferred treatments are forglioblastoma and osteoblastomas. Preferred leukemia and lymphomatreatments are for acute lymphoblastic leukemia (ALL), acute myelogenousleukemia (AML) and Hodgkin's disease.

The solid state forms of Compound 1 disclosed herein, e.g. an anhydrouscrystalline form such as Form III, are useful to treat, prevent, slowthe progression of, or ameliorate one or more symptoms of a cancer,precancer or other hyperproliferation conditions as described above.

In treating cancers or hyperproliferation conditions, the composition orformulation comprising a solid state form of Compound 1 may detectablymodulate, e.g., decrease or increase, the expression or level oractivity of one or more biomolecules associated with the prevention,establishment, maintenance or progression of the cancer orhyperproliferation condition. Such biomolecules include one or more ofcarcinoembryonic antigen, prostate specific antigen, her2/neu, Bcl-XL,bcl-2, p53, IL-1α, IL-1β, IL-6, or TNFα, GATA-3, COX-2, NFκB, IkB, anIkB kinase, e.g., IkB kinase-α, IkB kinase-β or IkB kinase-γ, NFAT,calcineurin, calmodulin, a ras protein such as H-ras or K-ras, cyclin D,cyclin E, xanthine oxidase, or their isoforms, homologs or mutant forms,which may have either reduced or enhanced biological activity(ies), andwhich may be detectably decreased. Biomolecules or their activity(ies)that can be detectably increased include IL-2, IFNγ, IL-12, T-bet,O⁶-methylguanine-DNA-methyltransferase, calcineurin, calmodulin, asuperoxide dismutase (e.g., Mn, Zn or Cu), a tumor suppressor proteinsuch as the retinoblastoma protein (Rb) or CDKN2A (p16), BRCA1, BRCA2,MeCP2, MBD2, PTEN, NBR1, NBR2 or the isoforms, homologs or mutant forms,which may have either attenuated or enhanced biological activity(ies),of any of these molecules. One or more of these biomolecules may bemodulated in any the cancers or conditions described herein.

In one embodiment, a decrease in circulating prostate specific antigen(PSA) or a decrease in velocity of PSA increase (e.g. decreased doublingtime in the increase of serum PSA levels) is the detectable changeconsistent with improvement in a subject having a hyperproliferationcondition wherein the condition is prostate cancer.

Dosing Protocols or Methods—

In treating any of the conditions or symptoms disclosed herein, one cancontinuously (daily) or intermittently administer the compositions orformulations comprising a crystalline form of Compound 1 to a subjectsuffering from or susceptible to the condition or symptom. In treating ahyperproliferation condition, such as prostate cancer, breast cancer orbenign prostate hyperplasia or other conditions disclosed herein with acomposition or formulation comprising a solid state form of Compound 1(e.g., crystalline Form I or crystalline Form III) intermittent dosingcan avoid or ameliorate some of the undesired aspects normallyassociated with discontinuous dosing. Such undesired aspects includefailure of the patient or subject to adhere to a daily dosing regimen,tendency to acquire disease tolerance to treatment or requirement toreduce the dosages of other therapeutic agents given concomitantly dueto their associated unwanted side effects or toxicities. Intermittentdosing is also employed if tachyphylaxis occurs whereupon the dosingschedule is adjusted to avoid or minimize the adverse response.

In any of the continuous (daily) or intermittent dosing protocolsdescribed herein, or in treating any of the diseases, conditions orsymptoms described herein, an appropriate composition or formulationcomprising a solid state form of Compound 1 (e.g. crystalline Form I orcrystalline Form III) can be administered by one or more suitableroutes, e.g., oral, buccal, sublingual, intramuscular (i.m.),subcutaneous (s.c.), intravenous (i.v.), intradermal, another parenteralroute or by an aerosol wherein the active pharmaceutical ingredient is asolid state form of Compound 1 (e.g. crystalline Form I or crystallineForm III). The daily dose in such methods may include about 0.0025mg/kg/day to about 5.0 mg/Kg/day. Typically, the daily dose in suchadministration methods will comprise about 0.01 mg/kg/day Compound 1 insolid state form (e.g., crystalline Form I or crystalline Form III), toabout 3 mg/kg/day, or about 0.1 to about 1 mg/kg/day, including about0.3 mg/kg/day to 0.5 mg/kg/day. Higher dosages, e.g., to about 60mg/Kg/day, may also be used in some veterinary applications. In someembodiments, suspension formulations described herein comprising acrystalline form of Compound 1 are administered i.m. or s.c. Preferredunit doses contain 0.5 to 100 mg or more of one or more, typically oneor two solid state forms of Compound 1 typically administered q.d,b.i.d, t.i.d or q.i.d. with 5, 15, 10 or 25 mg of a single solid stateform of Compound 1 administered b.i.d. particularly preferred. Preferredunit dosage forms include those suitable for oral dosing such as tabletsand capsules.

In some embodiments, such as treating a hormone sensitive or hormoneassociated cancer, a composition or formulation comprising a solid stateform of Compound 1 is administered daily q.d. or b.i.d for 14-180 days,typically 30-90 days or until a biomarker for the hyperproliferativecondition being treated indicates subsidence, lack of progression (i.e.stabilization) or remission of the disease. Dosing then resumes if thebiomarker indicates resurgence or recurrence of the disease. In one suchembodiment the cancer is prostate cancer and the biomarker iscirculating prostate specific antigen. In another such embodiment thecancer is metastatic prostate cancer and the biomarker is the spread ofdisease to the bone as estimated from bone scans. In another embodiment,the cancer is breast cancer and the biomarker is tumor burden.

Dosages of Compound 1 in solid state form administered by the routesdescribed herein and the use of combination therapies with otherstandard therapeutic agents or treatments could be applied essentiallyas described above for any of the diseases or conditions that aredisclosed herein. Thus, the Compound 1 in solid state form may beadministered prophylactically or therapeutically in chronic conditionsor they may be administered at the time of or relatively soon after anacute event such as a pain flare associated with a condition beingtreated.

Dosages of Compound 1 in solid state form, routes of administration andthe use of combination therapies with other standard therapeutic agentsor treatments could be applied essentially as described above for canceror hyperproliferation conditions or other conditions as disclosedherein. This, in some embodiments, the use of a solid state form ofCompound 1 is optionally combined with one or more additional therapiesfor a cancer or precancer(s), e.g., one or more of surgery and treatmentwith an antiandrogen or an antiestrogen as described herein or in thecited references, an antineoplastic agent such as an alkylating agent, anitrogen mustard, a nitrosourea, an antimetabolite or cytotoxic agent,or an analgesic such as propoxyphene napsylate, acetaminophen orcodeine. Exemplary anticancer and adjunct agents include methotrexate,thioguanine, mercaptopurine, adriamycin, chlorambucil, cyclophosphamide,cisplatin, procarbazine, hydroxyurea, allopurinol, erythropoietin,G-CSF, bicalutamide, anastrozole, fludarabine phosphate and doxorubicin.Such therapies would be used essentially according to standard protocolsand they would precede, be essentially concurrent with and/or followtreatment with a solid state form of Compound 1. In some embodiments,such additional therapies will be administered at the same time that asolid state form of Compound 1 is being used or within about 1 day toabout 16 weeks before or after at least one round of treatment with thesolid state form of Compound 1 is completed. Other exemplary therapeuticagents and their use have been described in detail, see, e.g.,Physicians Desk Reference 54^(th) edition, 2000, pages 303-3250, ISBN1-56363-330-2, Medical Economics Co., Inc., Montvale, N.J. One or moreof these exemplary agents can be used in combination with a solid stateform of Compound 1 to ameliorate, slow the establishment or progressionof, prevent or treat any of the appropriate cancers, precancers orrelated conditions described herein, or any of their symptoms.

In one embodiment the cancer being treated is prostate cancer which maybe androgen ablation sensitive or androgen ablation insensitive. If thecancer is androgen ablation sensitive, a solid state form of Compound 1is administered in an appropriate formulation either alone incombination with androgen receptor antagonist such as CASODEX™ (alsoknown as bicalutamide) optionally in combination with luteinizinghormone-releasing hormone or Leuprorelin (a gonadotropin-releasinghormone agonist), to a subject with prostate cancer who may or may notbe castrated. The combination therapies may be co-administered eithercombined within a single dosage form (e.g., co-formulation of Compound 1with bicalutamide) or in separate dosage forms (e.g. Compound 1formulated in an oral dosage form with bicalutamide in a separate oraldosage form or with a hormone agonist administered parenterally in asuspension formulation) either contemporaneously with each other (0-15min apart) or within 15 min-24 hours, 30 min-24 hours apart or 1-24hours apart, or administered using different dosage schedules (i.e.,administered on different days) to permit optimal additive orsynergistic interactions with each therapy component or to minimizeadverse events by dosage reduction of one or more of the therapycomponents. If the subject with prostate cancer is androgen ablationinsensitive, which includes prostate cancer that isandrogen-independent, castrate-independent or hormone refractory, asolid state form of Compound 1 in an appropriate formulation isadministered in combination with a cytotoxic agent such as ananti-microtubule agent, including but not limited to a taxane compoundsuch as docetaxel or paclitaxel. The combination therapies may beco-administered either combined within a single dosage form (i.e.,co-formulated where appropriate) or in separate dosage forms aspreviously described. Again, dosing amount and schedule may be varied toprovide optimal therapy. Combination therapies using cytotoxic agentsare typically initiated after the subject becomes symptomatic withmetastatic disease.

In another embodiment the cancer being treated is breast cancer byadministration of a solid state form of Compound 1 in an appropriateformulation either alone in combination with one or more othertherapeutic agents common to the treatment of breast cancer. In oneembodiment a solid dosage from of Compound 1 is used in combination witha gonadotropin-releasing hormone agonist such as leuprolide. In anotherembodiment the solid dosage from of Compound 1 may be used incombination with HERCEPTIN™ depending on the Her2/neu status of thetumor. In another embodiment the solid dosage form of Compound 1 is usedin combination with an irreversible aromatase inhibitor such asexemestane, which forms a covalent bond with the aromatase enzyme orinhibitors such as anastrozole, or letrozole, which inhibit thearomatase enzyme by reversible competition. In another embodiment thesolid dosage form of Compound 1 is used in combination a selectiveestrogen receptor modulator, such as tamoxifen or raloxifene, dependingon the estrogen receptor status of the tumor. The combination therapiesmay be co-administered either combined within a single dosage form(i.e., co-formulated where appropriate) or in separate dosage forms aspreviously described. Again, dosing amount and schedule may be varied toprovide optimal therapy. Compound 1 in solid state form is typicallygiven to a subject having breast cancer alone or concurrent with hormonemanipulation. Hormone manipulation is typically accomplished withTAMOXIFEN™ and an aromatase inhibitor such as ARIMIDEX™ (anastrozole)and is conducted after initial primary therapy, which may be surgery,chemotherapy, radiation or a combination thereof, to prevent recurrence.Compound 1 is this setting may replace tamoxifen or be administered inaddition to TAMOXIFEN™. When used concurrently with a chemotherapeuticagent, as is sometime done in late stage disease, Compound 1 isadministered in combination with a taxane compound, such as docetaxel orpaclitaxel, to prolong the time before the next course of chemotherapyor is used to reduce the amount of chemotherapeutic agent to be given soas to mitigate the agent's side effects or to prevent or slow theemergence of resistance to the agent.

Numbered Embodiments

Several aspects of the invention and related subject matter include thefollowing numbered embodiments.

1. A solid state form of 17α-ethynyl-5α-androstane-3α,17β-diol.

2. The solid-state form of embodiment 1 wherein the solid-state form isone or more crystalline forms of 17α-ethynyl-5α-androstane-3α,17β-diolsubstantially free of 17α-ethynyl-5α-androstane-3α,17β-diol in amorphousform.

3. The solid-state form of embodiment 1 wherein solid-state form is apolymorph or pseudopolymorph of 17α-ethynyl-5α-androstane-3α,17β-dioland is essentially free of amorphous17α-ethynyl-5α-androstane-3α,17β-diol.

4. The solid-state form of embodiment 1 wherein the solid-state form isa crystalline form of 17α-ethynyl-5α-androstane-3α,17β-diol and isessentially free of amorphous and other crystalline forms of17α-ethynyl-5α-androstane-3α,17β-diol.

5. The solid state form of embodiment 4 wherein the solid-state form ischaracterized by: (a) an X-ray powder pattern with 2-theta values of10.6, 14.7, 16.0±0.1 and optionally one or more 2-theta values of 12.3,14.3, 15.9, 16.4, 17.5, 20.3, 24.0 and 27.2±0.1 and (b) optionally withdifferential thermal analysis thermogram (DTA) having a broad endothermcentered at about 81° C. and a prominent endotherm at about 164° C.(onset at about 162° C.) obtained with a heating rate of 10° C./min oris characterized by (a) and (b).

6. The solid state form of embodiment 5 further characterized bythermogravimetric analysis (TGA) thermogram with 12 wt % weight lossfrom about 40° C. to about 105° C. obtained with a heating rate of 10°C./min.

7. The solid state form of embodiment 5 further characterized by DTAthermogram with an exotherm at about 100° C.

8. The solid state form of embodiment 4 wherein the solid-state form ischaracterized by: (a) an X-ray powder pattern with 2-theta values of11.3, 13.5, 16.2 and 16.5±0.1 and optionally one or more 2-theta valuesof 9.3, 9.9, 16.0. 17.4, and 19.0±0.10 and (b) optionally with DTAthermogram having a prominent endotherm at about 164° C. (onset at about162° C.) and a TGA thermogram having negligible wt % weight loss fromabout 40° C. to about 105° C., obtained with a heating rate of 10°C./min or is characterized by (a) and (b).

9. The solid-state form of embodiment 8 further characterized by Ramanspectra substantially identical to FIG. 6 or FIG. 7.

10. The solid-state form of embodiment 4 wherein the solid-state form ischaracterized by an X-ray powder diffraction pattern and differentialscanning calorimetry (DSC) thermogram substantially identical to theX-ray powder diffraction pattern of FIG. 9 and DSC-TGA thermograms ofFIG. 10.

11. The solid-state form of embodiment 10 further characterized by Ramanspectra substantially identical to FIG. 11 or FIG. 12.

12. The solid-state form of embodiment 4 wherein the solid-state form ischaracterized by an X-ray powder diffraction pattern and DSC-TGAthermograms substantially identical to the X-ray powder diffractionpattern of FIG. 1 or FIG. 2 and DSC-TGA thermograms of FIG. 3.

13. The solid state form of embodiment 3 wherein the solid-state form ischaracterized by: (a) an X-ray powder pattern with 2-theta values of9.8, 13.0, 14.7 and 17.0±0.1 and optionally one or more 2-theta valuesof 8.3, 11.3, 13.9, 15.0, 15.4, 16.1, 16.5, 17.8, 18.7, 20.0, 20.8, 22.1and 25.1±0.1 and (b) optionally with DTA thermogram having a broadendotherm centered at about 88 C and a prominent endotherm at about 164°C. (onset at about 162° C.). 14. The solid state form of embodiment 13further characterized by a TGA thermogram having between about 5-6 wt %weight loss from about 60° C. to about 105° C. or about 7 wt % weightloss from about 40° C. to about 160° C., obtained with a heating rate of10° C./min or is characterized by (a) and (b).

15. The solid state form of embodiment 13 further characterized by DTAthermogram with an exotherm at about 106° C.

16. The solid state form of embodiment 3 wherein the solid-state form ischaracterized by an X-ray powder diffraction pattern and DTA-TGAthermograms substantially identical to the X-ray powder diffractionpattern of FIG. 15 and DTA-TGA thermograms of FIG. 16.

17. The solid state form of embodiment 3 wherein the solid-state form ischaracterized by: (a) an X-ray powder pattern with 2-theta values of5.8, 9.5, 11.5, 15.2 and 18.9±0.1 and optionally one or more 2-thetavalues of 13.5, 16.0, 16.5, 17.3, 19.3, 20.9, 24.5 and 29.3±0.10 and (b)optionally with DTA thermogram having a prominent endotherm at about164° C. (onset at about 162° C.) and TGA thermogram with negligibleweight loss from about 40° C. to about 160° C., obtained with a heatingrate of 10° C./min.

18. The solid state form of embodiment 3 wherein the solid-state form ischaracterized by an X-ray powder diffraction pattern substantiallyidentical to the X-ray diffraction pattern of FIG. 18.

19. The solid state form of embodiment 3 wherein the solid-state form ischaracterized by: (a) an X-ray powder pattern with 2-theta values of9.8, 13.3, 15.0 and 18.7±0.1 and optionally one or more 2-theta valuesof 6.7, 7.2, 7.5, 14.3, 14.6, 16.0, 17.0, 17.7, 18.3, 20.9 and 21.8±0.1and (b) optionally with DTA thermogram having a prominent endotherm atabout 164° C. (onset at about 162° C.) and TGA thermogram with about 5wt % weight loss from about 40° C. to about 85° C. or about 12% weightloss from about 40° C. to about 180° C., obtained with a heating rate of10° C./min.

20. The solid state form of embodiment 3 wherein the solid-state form ischaracterized by an X-ray powder diffraction pattern substantiallyidentical to the X-ray diffraction pattern of FIG. 19 and DTA-TGAthermograms substantially identical to the DTA-TGA thermograms of FIG.20.

21. The solid state form of embodiment 3 wherein the solid-state form ischaracterized by: (a) an X-ray powder pattern with 2-theta values of9.8, 13.5, 14.2, 15.8, 19.5±0.1 and optionally one or more 2-thetavalues of 5.9, 8.3, 11.4, 11.9, 17.8, 21.4 and 26.9±0.10 and (b)optionally with DTA thermogram having a prominent endotherm at about164° C. (onset at about 162° C.) and TGA thermogram with negligle wt %weight loss from about 40° C. to about 160° C.

22. The solid state form of embodiment 3 wherein the solid-state form ischaracterized by an X-ray powder diffraction pattern substantiallyidentical to the X-ray diffraction pattern of FIG. 21.

23. The solid state form of embodiment 3 wherein the solid-state form ischaracterized by: (a) an X-ray powder pattern with 2-theta values of11.1, 16.0, 11.6, 17.7 and 18.7±0.1 and optionally one or more 2-thetavalues of 9.4, 10.1, 19.1, 23.7, 24.3 and 28.4±0.10 and (b) optionallywith DTA thermogram having a prominent endotherm at about 164° C. (onsetat about 162° C.) and TGA thermogram with negligle wt % weight loss fromabout 40° C. to about 160° C.

24. The solid state form of embodiment 3 wherein the solid-state form ischaracterized by an X-ray powder diffraction pattern substantiallyidentical to the X-ray diffraction pattern of FIG. 22.

25. The solid-state form of embodiment 1 wherein the solid-state form isamorphous 17α-ethynyl-5α-androstane-3α,17β-diol substantially free of17α-ethynyl-5α-androstane-3α,17β-diol in crystalline form.

26. The solid-state form of embodiment 1 wherein the solid-state form isamorphous 17α-ethynyl-5α-androstane-3α,17β-diol essentially free of17α-ethynyl-5α-androstane-3α,17β-diol in crystalline form

27. A solid formulation comprising a solid state form of17α-ethynyl-5α-androstane-3α,17β-diol and at least one pharmaceuticallyacceptable excipient.

28. The formulation of embodiment 27 wherein the solid state form is oneor more crystalline forms of 17α-ethynyl-5α-androstane-3α,17β-diol.

29. The formulation of embodiment 28 wherein one crystalline form is apolymorph or pseudopolymorph form of17α-ethynyl-5α-androstane-3α,17β-diol and is substantially free of17α-ethynyl-5α-androstane-3α,17β-diol in amorphous form.

30. The formulation of embodiment 28 wherein one crystalline form is apolymorph or pseudopolymorph form of17α-ethynyl-5α-androstane-3α,17β-diol and is essentially free ofamorphous 17α-ethynyl-5α-androstane-3α,17β-diol.

31. The formulation of embodiment 27 wherein the solid-state form is acrystalline form of 17α-ethynyl-5α-androstane-3α,17β-diol and isessentially free of amorphous and other crystalline forms of17α-ethynyl-5α-androstane-3α,17β-diol.

32. The formulation of embodiment 31 wherein the crystalline form is ananhydrate, optionally selected from the group consisting of crystallineForm III. Form V, Form VII and Form VIII

33. The formulation of embodiment 31 wherein the crystalline form is asolvate, optionally selected from the group consisting of crystallineForm I and Form IV

34. The formulation of embodiment 31 wherein the crystalline form isForm I

35. The formulation of embodiment 31 wherein the crystalline form isForm III.

36. The formulation of embodiment 31 wherein the crystalline form isForm IV

37. The formulation of embodiment 31 wherein the crystalline form isForm V.

38. The formulation of embodiment 31 wherein the crystalline form isForm VII.

39. The formulation of embodiment 31 wherein the crystalline form isForm VIII.

40. The formulation of embodiment 27 wherein the solid state form isamorphous 17α-ethynyl-5α-androstane-3α,17β-diol.

41. The formulation of embodiment 41 wherein amorphous17α-ethynyl-5α-androstane-3α,17β-diol is substantially free ofcrystalline 17α-ethynyl-5α-androstane-3α,17β-diol.

42. The formulation of any one of embodiments 27-41 wherein theformulation is in a capsule or tablet for oral dosing and thepharmaceutically acceptable excipient is a surface active agent in anamount sufficient to provide 90% dissolution of the formulation in waterat ambient temperature after 30 min.

43. The formulation of embodiment 42 wherein the surface active agent issodium lauryl sulfate.

44. The formulation of any one of embodiments 27-41 wherein thepharmaceutically acceptable excipients are comprised of sodium laurylsulfate, microcrystalline cellulose and magnesium stearate

45. The formulation of any one of embodiments 27-41 wherein thepharmaceutically acceptable excipients consist essentially of sodiumlauryl sulfate, microcrystalline cellulose and magnesium stearate inrelative amounts to the solid state form of17α-ethynyl-5α-androstane-3α,17β-triol as provided by Table 13 or Table14.

46. A method to treat a hyperproliferation condition comprisingadministering to a subject in need thereof an effective amount of17α-ethynyl-5α-androstane-3α,17β-triol in a solid state form or in asolid formulation comprising the solid state form of17α-ethynyl-5α-androstane-3α,17β-triol and at least one pharmaceuticallyacceptable excipient.

47. The method of embodiment 46 wherein the solid state form is acrystalline form of 17α-ethynyl-5α-androstane-3α,17β-triol substantiallyfree of 17α-ethynyl-5α-androstane-3α,17β-triol in amorphous form.

48. The method of embodiment 47 wherein the solid state form is apolymorph or pseudopolymorph form of17α-ethynyl-5α-androstane-3α,17β-triol essentially free or substantiallyfree of amorphous and other crystalline forms of17α-ethynyl-5α-androstane-3α,17β-triol.

49. The method of embodiment 48 wherein the polymorph or pseudopolymorphform is crystalline Form III.

50. The method of embodiment 46 wherein the hyperproliferation conditionis a hormone sensitive or hormone associated cancer.

51. The method of embodiment 46 wherein the hyperproliferation conditionis prostate cancer, breast cancer, benign prostatic hyperplasia orprostatic interstitial neoplasia.

52. A method of preparing a solid formulation of any one in embodiments12-24 comprising the step of blending a solid state form of17α-ethynyl-5α-androstane-3α,17β-triol with one, two, three or fourpharmaceutically acceptable excipients wherein at least one excipient isa surface active agent.

53. The method of embodiment 52 wherein the solid state form iscrystalline Form III.

54. The method of embodiment 52 wherein the solid state form isamorphous 17α-ethynyl-5α-androstane-3α,17β-triol.

55. The method of embodiment 52 wherein at least one excipient is sodiumlauryl sulfate.

56. A method of preparing a liquid formulation comprising17α-ethynyl-5α-androstane-3α,17β-triol and a pharmaceutically acceptableexcipients wherein at least one excipient is a liquid excipientcomprising the step of contacting or admixing a solid state form of17α-ethynyl-5α-androstane-3α,17β-triol with the liquid excipient,optionally in the presence of another excipient.

57. The method of embodiment 56 wherein the solid state form iscrystalline Form III.

58. The method of embodiment 56 wherein the solid state form isamorphous 17α-ethynyl-5α-androstane-3α,17β-triol.

59. A method to treat a hyperproliferation condition comprisingadministering to a subject in need thereof an effective amount of17α-ethynyl-5α-androstane-3α,17β-triol in a liquid formulation preparedaccording to the method of claim 57, 58 or 59.

60. The method of embodiment 59 wherein the hyperproliferation conditionis a hormone associated or a hormone sensitive cancer.

61. The method of embodiment 60 wherein the hyperproliferation conditionprostate cancer, breast cancer, benign prostatic hyperplasia orprostatic interstitial neoplasia.

62. A product prepared by a process comprising the steps of admixing amethanolic solution of Compound 1 with sufficient water to form aprecipitate.

63. A product prepared by a process comprising the step of heating apseudopolymorph of Compound 1 under reduced pressure to effectdesolvation of the pseudopolymorph.

64. The product of embodiment 63 wherein the peudopolymorph is Form I

65. A product prepared by a process comprising the steps of admixing ahot EtOAc solution of Compound 1 with sufficient heptane to effectcrystallization upon cooling to ambient temperature.

66. A product prepared by a process comprising the steps of (1) removingsolvent from a THF solution of Compound 1 under ambient temperature andpressure to provide a gel (2) removing residual solvent from the gelunder reduced pressure at ambient temperature.

67. A product prepared by a process comprising the step of fastevaporation of a solution of Compound 1 in 2:3 acetonitile:water oradmixing an acetonitrile solution of Compound 1 with sufficient water toeffect crash precipitation.

68. A product prepared by a process comprising the steps of (1) admixingan acetonitrile solution of Compound 1 with water to provide 2:3acetonitile:water solution and (2) removing solvent from theacetonitrile-water solution under ambient temperature and pressure.

69. A product prepared by a process comprising the steps of (3) removingsolvent from an acetone solution of Compound 1 under ambient temperatureand pressure.

70. A product prepared by a process comprising the steps of (1) admixingForm III Compound 1 with 2,2,2-trifluoroethanol (TFE) to form a slurry(2) agitating the TFE slurry at about 30° C. for up to about 6 days.

71. A product prepared by a process comprising the steps of (1) removingsolvent from a dioxane solution of Compound 1 under ambient temperatureand pressure to provide a gel (2) removing residual solvent from the gelunder reduced pressure for up to 1 day at ambient temperature.

72. A product prepared by a process comprising the steps of (1) admixinga ethanolic slurry of Compound 1 with sufficient isopropyl acetate toeffect dissolution under sonication; (2) removing sufficient solventfrom the ethanol-isopropyl acetate solution to form solids such thatcomplete re-dissolution will occur upon heating to about 47° C.; (3)cooling the re-dissolved solution to about 5° C.

73. A product prepared by a process comprising the steps of removingsolvent from a dichloromethane solution of Compound 1 under ambienttemperature and pressure.

Further aspects of the invention related to crystalline17α-ethynyl-androstane-3α,17β-diol includes the following numberedembodiments.

1A. A crystalline form 17α-ethynyl-5α-androstane-3α,17β-diol.

2A. The crystalline form of embodiment 1A wherein the crystalline formis a pseudopolymorph, a polymorph or a mixture thereof.

3A. The crystalline form of embodiment 2A wherein the pseudopolymorph isa solvate.

4A. The crystalline form of embodiment 2A wherein the pseudopolymorph isa hydrate.

5A. The crystalline form of embodiment 3A wherein the solvate is a mixedsolvate of water and an alcohol.

6A. The crystalline form of claim 2A wherein the crystalline form is apseudopolymorph wherein the pseudopolymorph consists essentially of17α-ethynyl-5α-androstane-3α,17β-diol, water and an alcohol wherein thealcohol is methanol.

7A. The crystalline form of 4A wherein the pseudopolymorph is a mixedsolvate containing water and methanol in a water:methanol ratio ofbetween about 2:1 to 1:1.

8A. The crystalline form of embodiment 4A wherein the crystalline formis a pseudopolymorph characterized by the molecular formula ofC₂₁H₃₂O₂.1CH₃OH.1H₂O.

9A. The crystalline form of embodiment 4A wherein the pseudopolymorph isessentially free of 17α-ethynyl-5α-androstane-3α,17β-diol in othercrystalline forms and has a thermal gravimetric analysis thermogram withabout 12 wt % when heated from about 25° C. to 60° C. to about 105° C.using a temperature ramp of 10° C./min.

10A. The crystalline form of embodiment 1A wherein the crystalline formis a product prepared by a process comprising the step of partial orcomplete desolvation of a pseudopolymorph of17α-ethynyl-5α-androstane-3α,17β-diol.

11A. The crystalline form of embodiment 10A wherein the pseudopolymorphis a hydrate or a mixed solvate of water and methanol.

12A. The crystalline form of embodiment 10A wherein the pseudopolymorphis crystalline Form I, Form IV or Form VI

13A. The crystalline form of embodiment 1A wherein the crystalline formis an anhydrate.

14A. The crystalline form of embodiment 13A wherein the anhydrate is aproduct prepared from a process comprising the step of completedesolvation of crystalline Form I, Form IV or Form VI.

15A. The crystalline form of embodiment 13A wherein the anhydrate iscrystalline Form III.

16A. The crystalline form of embodiment 1A wherein the crystalline formis characterized by one or more, typically 2, 3 or 4 XRPD prominentpeaks in Table 1 or Table 2 optionally with a thermogram event obtainedfrom a thermal analysis method disclosed herein.

17A. The crystalline form of embodiment 1A wherein the crystalline formis characterized by one or more, typically 2, 3 or 4 XRPD prominentpeaks in Table 5. optionally with a thermogram event obtained from athermal analysis method disclosed herein.

18A. The crystalline form of embodiment 1A wherein the crystalline formis characterized by one or more, typically 2, 3 or 4 XRPD prominentpeaks in Table 8. optionally with a thermogram event obtained from athermal analysis method disclosed herein.

19A. The crystalline form of embodiment 1A wherein the crystalline formis characterized by one or more, typically 2, 3 or 4 XRPD prominentpeaks in Table 9. optionally with a thermogram event obtained from athermal analysis method disclosed herein.

20A. The crystalline form of embodiment 1A wherein the crystalline formis characterized by one or more, typically 2, 3 or 4 XRPD prominentpeaks in Table 10. optionally with a thermogram event obtained from athermal analysis method disclosed herein.

21A. The crystalline form of embodiment 1A wherein the crystalline formis characterized by one or more, typically 2, 3 or 4 XRPD prominentpeaks in Table 11. optionally with a thermogram event obtained from athermal analysis method disclosed herein.

22A. The crystalline form of embodiment 1A wherein the crystalline formis characterized by one or more, typically 2, 3 or 4 XRPD prominentpeaks in Table 12. optionally with a thermogram event obtained from athermal analysis method disclosed herein.

23A. The crystalline form of embodiment 1A wherein the crystalline formis characterized by a pairwise distribution function calculated from theXRPD pattern of FIG. 2.

24A. The crystalline form of embodiment 1A wherein the crystalline formis characterized by a pairwise distribution function calculated from theXRPD pattern of FIG. 9.

25A. The crystalline form of embodiment 1A wherein the crystalline formis characterized by one or more absorptions, typically one or twoabsorptions, in Raman identified in FIG. 6 or FIG. 7.

26A. The crystalline form of embodiment 1A wherein the crystalline formis characterized by one or more absorptions, typically one or twoabsorptions, in Raman identified in FIG. 11 or FIG. 12.

Further aspects of the invention related to crystalline17α-ethynyl-androstane-3α,17β-diol includes the following numberedembodiments.

1B. Crystalline 17α-ethynyl-androstane-3α,17β-diol.

2B. The crystalline 17α-ethynyl-androstane-3α,17β-diol of embodiment 1Bwherein the crystalline 17α-ethynyl-androstane-3α,17β-diol issubstantially free of amorphous 17α-ethynyl-androstane-3α,17β-diol,optionally as characterized an analytic method described herein such asXRPD, DSC, TGA, melting point, Raman spectroscopy, Karl Fisher and/orelemental analysis. Crystalline forms of17α-ethynyl-androstane-3α,17β-diol includes anhydrates, hydrates andsolvates, which include mixed water-solvent solvates. In theseembodiments, crystalline 17α-ethynyl-androstane-3α,17β-diol that issubstantially free of amorphous 17α-ethynyl-androstane-3α,17β-diol willtypically and preferably contain less than about 10% w/w or less thanabout 7% w/w of the amorphous material.

3B. The crystalline 17α-ethynyl-androstane-3α,17β-diol of embodiment 1Bor 2B as Form I crystals. This form of17α-ethynyl-androstane-3α,17β-diol is a mixed solvate material with a1:1:1 ratio of 17α-ethynyl-androstane-3α,17β-diol:water:methanol and istypically substantially free of amorphous17α-ethynyl-androstane-3α,17β-diol.

4B. The crystalline Form I 17α-ethynyl-androstane-3α,17β-diol ofembodiment 3B that contains less than about 10% w/w or less than about7% w/w of other crystalline forms of 17α-ethynyl-androstane-3α,17β-diol,optionally as characterized by an analytic method described herein suchas XRPD, DSC, TGA, melting point, Raman spectroscopy, Karl Fisher and/orelemental analysis.

5B. The crystalline of 17α-ethynyl-androstane-3α,17β-diol of embodiment3A characterized by (1) space group P2₁2₁2₁ (No. 19); Z=4 or (2) unitcell parameters of a=7.4893 (4) Å, b=11.0586 (8) Å, c=25.5095 (15) Å,α=90.00°, β=90.00°, γ=90.00°, V=2112.7 (2) Å³.

6B. The crystalline form of 17α-ethynyl-androstane-3α,17β-diol ofembodiment 3A characterized by (1) an XRPD pattern with prominent peaksat about 10.59, 12.33, 14.29, 14.72, 16.04, 16.41, 17.49, 20.27, 24.04and 27.21 degrees 2θ, optionally with (2) a DTA or DSC thermogram havingan endotherm with onset at about 160° C.±3° and TGA thermogram with wt %loss of at least about 10% or water content of about 5% by Karl-Fischertitration

7B. The crystalline form or 17α-ethynyl-androstane-3α,17β-diol ofembodiment 3A characterized by (1) an XRPD pattern with peaks at about9.30, 9.85, 11.33, 13.45, 15.96, 16.16, 16.48 and 17.42 degrees 2θoptionally with (2) a DTA or DSC thermogram having an endotherm onset atabout 160° C.±3° and a TGA thermogram with negligible wt % loss, weightof 2% or less, or weight loss of 0.3% from about 40° C. to about 105° C.or from about 40° C. to about 160° C. using a temperature ramp of 10°C./min.

8B. The crystalline form of embodiment 1B or 2B wherein the crystallineis Form III characterized by sufficient bioavailability of thecrystalline material to be suitable for once daily or twice dailyadministration of unit oral doses of 5 mg, 10 mg, 15 mg, 20 mg or 50 mgto a human, such as a human having a cancer or a precancer, optionallybenign prostatic hypertrophy, prostate cancer or breast cancer.

9B. The crystalline form of embodiment 1B, 2B or 8B wherein thecrystalline form is Form III characterized by sufficient stability onstorage at 65° C. and 75% relative humidity for at least 6 monthswherein sufficient stability is characterized by a change of less thanabout 5% w/w in the degradation of 17α-ethynyl-androstane-3α,17β-diol toa degradant or by conversion of less than about 5% w/w to another solidstate form.

10B. The crystalline 17α-ethynyl-androstane-3α,17β-diol of embodiment 1Bor 2B as Form III crystals. This form of17α-ethynyl-androstane-3α,17β-diol is an anhydrate and does not containa solvent as measured by an analytical method described herein such asKarl Fisher titration, gas chromatography analysis, proton-NMRspectroscopy and/or elemental analysis, and in preferred embodiments itis substantially free of amorphous 17α-ethynyl-androstane-3α,17β-diol,optionally as measured by an analytical method described herein such asXRPD, DSC/DTA, TGA, Raman spectroscopy or solid state NMR spectroscopy

11B. The crystalline Form III 17α-ethynyl-androstane-3α,17β-diol ofembodiment 10B that contains less than about 10% w/w or less than about7% w/w of other crystalline forms of 17α-ethynyl-androstane-3α,17β-diol,optionally as characterized by an analytical method described hereinsuch as XRPD, DSC/DTA, TGA, Raman spectroscopy or solid state NMRspectroscopy.

12B. The crystalline Form III 17α-ethynyl-androstane-3α,17β-diol ofembodiment 10B or 11B having (1) an XRPD pattern with prominent peaks atabout 9.30, 9.85, 11.33, 13.45, 15.96, 16.16, 16.48 and 17.42 degrees or(2) a Raman trace substantially identical to that shown in FIG. 6 orFIG. 7 or a combination of (1) and (2).

The crystalline Form III 17α-ethynyl-androstane-3α,17β-diol ofembodiment 10B that contains less than about 10% w/w or less than about7% w/w of other crystalline forms of 17α-ethynyl-androstane-3α,17β-diol,optionally as characterized by an analytical method described hereinsuch as XRPD, DSC/DTA, TGA, Raman spectroscopy or solid state NMRspectroscopy.

12B. The crystalline Form I 17α-ethynyl-androstane-3α,17β-diol ofembodiment 3B characterized by (1) an XRPD pattern with prominent peaksat about 10.59, 12.33, 14.29, 14.72, 16.04, 16.41, 17.49, 20.27, 24.04and 27.21 degrees 2θ or (2) a Raman trace substantially identical tothat shown in FIG. 11 or FIG. 12 or a combination of (1) and (2).

13B. The crystalline 17α-ethynyl-androstane-3α,17β-diol of embodiment 1Bor 2B as Form IV crystals. This form of17α-ethynyl-androstane-3α,17β-diol is a hydrate and contains water in a1:1 ratio. In preferred embodiments it is substantially free ofamorphous 17α-ethynyl-androstane-3α,17β-diol, optionally as measured byan analytical method described herein such as XRPD, DSC/DTA, TGA, Ramanspectroscopy and/or solid state NMR.

14B. The crystalline Form IV 17α-ethynyl-androstane-3α,17β-diol ofembodiment 13B that contains less than about 10% w/w or less than about7% w/w of other crystalline forms of 17α-ethynyl-androstane-3α,17β-diol,optionally as characterized by an analytic method described herein suchas XRPD, DSC, TGA, Raman spectroscopy and/or solid state NMR.

15B. The crystalline Form IV 17α-ethynyl-androstane-3α,17β-diol ofembodiment 13B or 14B characterized by (1) an XRPD pattern withprominent peaks at about 8.31, 9.84, 11.28, 13.02, 13.86, 14.73, 15.00,16.14, 16.53, 17.01, 17.76 and 18.72 degrees 2θ, (2) a DTA or DSCthermogram with an exotherm onset at about 105° C.±3°, (3) a TGAthermogram with a weight loss on heating to the melting point thatcorresponds to complete loss of water for a monohydrate or (4) acombination of the foregoing such as (1) and (2), (1) and (3) or (2) and(3).

16B. The crystalline 17α-ethynyl-androstane-3α,17β-diol of embodiment 1Bor 2B as Form V crystals. This form of17α-ethynyl-androstane-3α,17β-diol is an anhydrate. In preferredembodiments it is substantially free of amorphous17α-ethynyl-androstane-3α,17β-diol, optionally as measured by ananalytical method described herein such as XRPD, DSC/DTA, TGA, Ramanspectroscopy and/or solid state NMR.

17B. The crystalline Form V 17α-ethynyl-androstane-3α,17β-diol ofembodiment 13B that contains less than about 10% w/w or less than about7% w/w of other crystalline forms of 17α-ethynyl-androstane-3α,17β-diol,optionally as characterized by an analytic method described herein suchas XRPD, DSC, TGA, melting point, Raman spectroscopy and/or solid stateNMR.

17B. The crystalline Form V 17α-ethynyl-androstane-3α,17β-diol ofembodiment 16B or 17B characterized by (1) an XRPD pattern withprominent peaks at about 5.82, 9.48, 11.49, 13.50, 15.21, 17.28 and18.93 degrees 2θ, (2) a TGA thermogram with negligible weight loss, 2%or less weight loss, or 0.3% or less weight loss on heating to themelting point (3) a DTA or DSC thermogram having an endotherm at about164° C. or (4) a combination of (1) and (2) or (1) and (3).

18B. The crystalline 17α-ethynyl-androstane-3α,17β-diol of embodiment 1Bor 2B as Form VI crystals. This form of17α-ethynyl-androstane-3α,17β-diol is a dioxane solvate in a 1:1 ratio.In preferred embodiments it is substantially free of amorphous17α-ethynyl-androstane-3α,17β-diol, optionally as measured by ananalytical method described herein such as XRPD, DSC/DTA, TGA, Ramanspectroscopy and/or solid state NMR.

19B. The crystalline Form VI 17α-ethynyl-androstane-3α,17β-diol ofembodiment 18B that contains less than about 10% w/w or less than about7% w/w of other crystalline forms of 17α-ethynyl-androstane-3α,17β-diol,optionally as characterized by an analytical method described hereinsuch as XRPD, DSC/DTA, TGA, Raman spectroscopy and/or solid state NMR.

20B. The crystalline Form VI 17α-ethynyl-androstane-3α,17β-diol ofembodiment 16B or 17B characterized by (1) an XRPD pattern withprominent peaks at about 7.17, 9.78, 13.26, 14.25, 14.61, 15.00 and18.69 degrees 2θ, (2) a TGA thermogram with a weight loss on heating tothe melting point with the loss corresponding to complete loss ofdioxane from a mono solvate (3) proton NMR spectrum obtained in CDCl₃with a peak at about δ=3.6 ppm, (4) a DSC or DTA thermogram with anendotherm at about 164° C. or (5) a combination of the forgoing such as(1) and (2), (1) and (3), (1) and (4) or (2), (3) and (4).

21B. The crystalline 17α-ethynyl-androstane-3α,17β-diol of embodiment 1Bor 2B as Form VII crystals. This form of17α-ethynyl-androstane-3α,17β-diol is an anhydrate. In preferredembodiments it is substantially free of amorphous17α-ethynyl-androstane-3α,17β-diol, optionally as measured by ananalytical method described herein such as XRPD, DSC/DTA, TGA, Ramanspectroscopy, and/or solid state NMR.

22B. The crystalline Form VII 17α-ethynyl-androstane-3α,17β-diol ofembodiment 19A that contains less than about 10% w/w or less than about7% w/w of other crystalline forms of 17α-ethynyl-androstane-3α,17β-diol,optionally as characterized by an analytical method described hereinsuch as XRPD, DSC/DTA, TGA, Raman spectroscopy and/or solid state NMR.

23B. The crystalline Form VII 17α-ethynyl-androstane-3α,17β-diol ofembodiment 20B or 21B characterized by (1) an XRPD pattern withprominent peaks at about 5.91, 9.78, 13.47, 14.16, 15.78, 17.85, 19.50and 21.45 degrees 2θ or (2) a TGA thermogram with negligible weightloss, 2% or less weight loss, or 0.3% or less weight loss on heating tothe melting point (3) a DTA or DSC thermogram having an endotherm atabout 164° C. or a combination of the foregoing such as (1) and (2), (1)and (3) or (1) (2) and (3).

24B. The crystalline 17α-ethynyl-androstane-3α,17β-diol of embodiment 1Bor 2B as Form VIII crystals. This form of17α-ethynyl-androstane-3α,17β-diol is an anhydrate. In preferredembodiments it is substantially free of amorphous17α-ethynyl-androstane-3α,17β-diol, optionally as measured by ananalytical method described herein such as XRPD, DSC/DTA, TGA, Ramanspectroscopy, and/or solid state NMR.

24B. The crystalline Form VIII 17α-ethynyl-androstane-3α,17β-diol ofembodiment 24B that contains less than about 10% w/w or less than about7% w/w of other crystalline forms of 17α-ethynyl-androstane-3α,17β-diol,optionally as characterized by an analytic method described herein suchas XRPD, DSC, TGA, melting point, Raman spectroscopy, Karl Fisher and/orelemental analysis.

25B. The crystalline Form VIII 17α-ethynyl-androstane-3α,17β-diol ofembodiment 23B or 24B characterized by (1) an XRPD pattern withprominent peaks at about 11.13, 15.96, 16.62, 17.76 and 18.75 degrees2θ, (2) a TGA thermogram with negligible weight loss, 2% or less weightloss, or 0.3% or less weight loss on heating to the melting point (3) aDTA or DSC thermogram having an endotherm at about 164° C. or acombination of the foregoing such as (1) and (2), (1) and (3) or (1) (2)and (3).

26B. Use of crystalline 17α-ethynyl-androstane-3α,17β-diol, or use of acomposition comprising one or more excipients and crystalline17α-ethynyl-androstane-3α,17β-diol, for the preparation of a medicamentfor the treatment or prophylaxis of a cancer or a precancer, optionallywherein the cancer or precancer is prostate cancer, breast cancer,ovarian cancer, endometrial cancer, lung cancer, pancreatic cancer orbenign prostatic hypertrophy. In these embodiments, the use ofcrystalline Forms I, III, IV, V, VII and VIII of17α-ethynyl-androstane-3α,17β-diol are preferred, with Forms III mostpreferred. In these uses appreciable amounts of two crystal forms can bepresent, but there is preferably only 1 crystalline form present, e.g.,a single crystal form comprises at least about 90% w/w or at least about93% w/w of the ethynyl-androstane-3α,17β-diol that is present.

27B. Use according to embodiment 26B wherein the crystalline17α-ethynyl-androstane-3α,17β-diol is substantially free of amorphous17α-ethynyl-androstane-3α,17β-diol, optionally as characterized by ananalytical method described herein such as XRPD, DSC/DTA, TGA, Ramanspectroscopy and/or solid state NMR.

Further aspects of the invention related to amorphous17α-ethynyl-androstane-3α,17β-diol include the following numberedembodiments.

1C. Amorphous 17α-ethynyl-5α-androstane-3α,17β-diol.

2C. The amorphous 17α-ethynyl-5α-androstane-3α,17β-diol of embodiment 1Cwherein the amorphous 17α-ethynyl-5α-androstane-3α,17β-diol issubstantially free of crystalline 17α-ethynyl-5α-androstane-3α,17β-diolas measured by XRPD analysis, optionally wherein the amorphous17α-ethynyl-5α-androstane-3α,17β-diol is substantially free ofcrystalline Form I and/or Form III17α-ethynyl-5α-androstane-3α,17β-diol.

3C. The amorphous 17α-ethynyl-5α-androstane-3α,17β-diol of embodiment 1Cor 2C wherein the amorphous 17α-ethynyl-5α-androstane-3α,17β-diolcontains less than about 8% w/w of crystalline17α-ethynyl-5α-androstane-3α,17β-diol.

4C. The amorphous 17α-ethynyl-androstane-3α,17β-diol of embodiment 1C,2C or 3C wherein the amorphous 17α-ethynyl-androstane-3α,17β-diolcontains less than about 5% w/w of crystalline17α-ethynyl-5α-androstane-3α,17β-diol.

5C. A pharmaceutical formulation comprising one or more excipients andamorphous 17α-ethynyl-5α-androstane-3α,17β-diol, optionally wherein theamorphous 17α-ethynyl-5α-androstane-3α,17β-diol is as described inembodiment 1C, 2C, 3C or 4C.

6C. A product, amorphous 17α-ethynyl-5α-androstane-3α,17β-diol, producedby a process comprising the step of adding 10% by volume water to afiltered solution of about 0.25 mg/mL Compound 1 in methanol withagitation

7C. The product of embodiment 6C wherein the amorphous17α-ethynyl-5α-androstane-3α,17β-diol (1) is substantially free ofcrystalline 17α-ethynyl-androstane-3α,17β-diol as measured by XRPDanalysis, or (2) contains less than about 8% w/w of crystalline17α-ethynyl-5α-androstane-3α,17β-diol, or (3) contains less than about5% w/w of crystalline 17α-ethynyl-5α-androstane-3α,17β-diol, optionallywherein the crystalline 17α-ethynyl-5α-androstane-3α,17β-diol is Form Iand/or Form III 17α-ethynyl-5α-androstane-3α,17β-diol.

8C. Use of amorphous 17α-ethynyl-5α-androstane-3α,17β-diol, or use of acomposition comprising one or more excipients and amorphous17α-ethynyl-5α-androstane-3α,17β-diol for the preparation of amedicament for the treatment or prophylaxis of a cancer, precancer orhyperplasia, optionally wherein the cancer or hyperplasia is prostatecancer, breast cancer, ovarian cancer, endometrial cancer or benignprostatic hypertrophy. In these uses, amorphous material preferablycomprises at least about 90% w/w or at least about 95% w/w of the17α-ethynyl-5α-androstane-3α,17β-diol that is present.

9C. Use according to embodiment 8C wherein the amorphous17α-ethynyl-androstane-3α,17β-diol is substantially free of crystalline17α-ethynyl-5α-androstane-3α,17β-diol as measured by XRPD analysis orwherein the amorphous 17α-ethynyl-5α-androstane-3α,17β-diol containsless than about 8% w/w or less than about 5% w/w of crystalline17α-ethynyl-5α-androstane-3α,17β-diol.

EXAMPLES Example 1 Synthesis of 17-ethynyl-5α-androstane-3α,17β-diol

Step A. Synthesis of 3α-trimethylsilyoxy-androst-5-en-17-one(TMS-3α-DHEA): 3α-DHEA is combined with 1,1,1,3,3,3-hexamethyldisilazane(HMDS) and saccharin (as catalyst) in acetonitrile. The reaction mixtureis heated to reflux for several hours with stirring under a nitrogenatmosphere. Liberated ammonia is purged under slight vacuum. The volumeis then reduced by distillation, followed by cooling the mixture andcollecting the precipitated product by filtration. The filter cake ofTMS-3α-DHEA product is washed with cold acetonitrile and dried with warmnitrogen to provide the title compound.

Step B. n-Butyl lithium is added slowly to Me₃Si—C≡CH in THF under anitrogen atmosphere at approximately 0° C. to produce the lithiumacetylide Me₃Si—C≡C—Li. The temperature is raised to about 20° C., andTMS-3α-DHEA is added as a solution in THF, and stirred for about 3hours. The reaction is quenched by raising the temperature to about 40°C. followed by the slow addition of methanol. Liberated acetylene ispurged under slight vacuum. Concentrated KOH is then slowly added untilgas evolution subsides, and the volume is reduced by approximately 50%by vacuum distillation at approximately 45° C. Excess 6 N HCl is slowlyadded, while maintaining the temperature at approximately 40° C. Thereaction mixture is diluted with water and chilled to approximately 5°C. before collecting the product by filtration and washing the filtercake with cold 50/50 methanol water. The product is dried with warmnitrogen to provide 17β-ethynyl-androst-5-ene-3α,17α-diol.

Step C. To 9.0 Kg of the title compound in a 250 L reactor was added71.2 Kg methanol. The agitated mixture was heated to reflux until thesolids had dissolved. After cooling to 55-60° C. the reactor contentswere filtered through a 25-micron filter and the reactor was then rinsedwith 2.4 Kg MeOH heated to 55-60° C. and filtered as above to combinethe filtrates. To the combined filtrates agitated in a 250 L reactor wasadded over a period of 30-60 min. 81.0 Kg of deionized water to form aslurry while maintaining the temperature between 35-60° C. The slurrywas then cooled to 0-5° C. over a period of at least 2 h and thetemperature was maintained for at least 1 h whereupon the solids werecollected by filtration. The filter cake was washed by slurring with 10Kg deionized water (repeated 2×). The filter cake was allowed to dryunder vacuum at about 28.5 in. Hg at about 45° C. to loss on drying of0.5% or less. Obtained was 8.4 Kg of the title material in crystallineform.

The crystalline form of Compound 1 obtained from this syntheticprocedure is represented by the low resolution XRPD pattern of FIG. 1.Peak listing for the X-Ray Powder Diffraction (XRPD) pattern of FIG. 1is provided in Table 1.

TABLE 1 Peak Listing for XRPD Pattern of Synthesis Product-LowResolution °2θ Intensity (%)  9.38 ± 0.10 9  9.66 ± 0.10 21 11.32 ± 0.1030 13.44 ± 0.10 100 15.94 ± 0.10 57 16.46 ± 0.10 37 17.38 ± 0.10 2118.66 ± 0.10 17 19.40 ± 0.10 40 19.44 ± 0.10 40 20.74 ± 0.10 10 21.44 ±0.10 11 21.56 ± 0.10 13 24.76 ± 0.10 10 25.42 ± 0.10 8 26.64 ± 0.10 1227.04 ± 0.10 8 27.68 ± 0.10 13 27.92 ± 0.10 10 28.94 ± 0.10 6 29.68 ±0.10 7 33.26 ± 0.10 6 34.00 ± 0.10 10 35.40 ± 0.10 10 36.44 ± 0.10 837.84 ± 0.10 10 38.64 ± 0.10 7

Thermal analysis for the synthesis product provides a thermogravimetricanalysis (TGA) thermogram showing negligible weight loss from about 35°C. to about 105° C. using a temperature ramp of 10° C./min and adifferential thermal analysis (DTA) thermogram showing a prominentendotherm at about 164° C. and is otherwise featureless. Thus, thermalanalysis indicates that the prominent or sole polymorphic form withinthe product is most likely an anhydrate. XRPD, DTA and TGA data is thusconsistent with crystalline Form III, discussed in subsequent example,as the dominant or sole crystalline form in the synthesis product.

Example 2

Preparation and Analysis of Crystalline Form III17α-ethynyl-5α-androstane-3α,17β-diol (Form III Compound 1): Compound 1(65.8 g) synthesis product from the immediately preceding example wasdried under vacuum at 80° C. to constant weight, typically requiringheating overnight, to provide 57.9 g. This dried material was dissolvedinto 230 mL HPLC grade EtOAc at 80° C. agitated by swirling. Thesolution was hot filtered through a coarse glass fritted funnel toremove residual solids then transferred to a 2 L flask to resume heatingat 80° C. HPLC grade heptane (1.158 L) was the added in about 100 mLaliquots with swirling. After completion of addition, the mixture wasallowed to cool to ambient temperature and allowed to stand overnight.The crystalline material was collected by filtration, air dried for 20min and under vacuum overnight to provide 48.8 g of Form III17α-ethynyl-5α-androstane-3α,17β-diol. Form III was also prepared bydissolving Compound 1 (44.0 mg) in tetrahydrofuran (600 μL). Thesolution was filtered through 0.2 μm nylon filter to a clean vial. Thesolvent was slowly evaporated under ambient conditions yielding a gelthat was vacuum dried at ambient temperature for 4 days to provideCompound 1 in crystalline Form III.

Additionally, Form III may produced by vapor stressing a melt-quenchsample of Compound 1 as described in a subsequent example for amorphousmaterial.

The high resolution XRPD for Form III, prepared according to theimmediately preceding procedure is given in FIG. 2. Peak listing forthis XRPD pattern is given in Table 2. Prominent peaks are at 9.30,9.85, 11.33, 13.45, 15.96, 16.16, 16.48 and 17.42±0.10 degrees 2θ.

TABLE 2 Peak Listing for XRPD Pattern of Form III-High Resolution °2θ dspace (Å) Intensity (%)  9.30 ± 0.10 9.506 ± 0.103 18  9.70 ± 0.10 9.122± 0.095 8  9.85 ± 0.10 8.976 ± 0.092 15 11.33 ± 0.10 7.807 ± 0.069 5313.45 ± 0.10 6.584 ± 0.049 100 15.96 ± 0.10 5.552 ± 0.035 29 16.16 ±0.10 5.484 ± 0.034 39 16.48 ± 0.10 5.379 ± 0.033 91 17.42 ± 0.10 5.092 ±0.029 24 18.70 ± 0.10 4.744 ± 0.025 5 18.95 ± 0.10 4.682 ± 0.025 1119.46 ± 0.10 4.563 ± 0.023 8 19.78 ± 0.10 4.488 ± 0.023 1 20.75 ± 0.104.281 ± 0.021 7 21.08 ± 0.10 4.214 ± 0.020 2 21.60 ± 0.10 4.114 ± 0.0192 22.79 ± 0.10 3.902 ± 0.017 2 24.62 ± 0.10 3.616 ± 0.015 1 24.80 ± 0.103.590 ± 0.014 6 25.01 ± 0.10 3.560 ± 0.014 8 25.39 ± 0.10 3.508 ± 0.0143 25.65 ± 0.10 3.474 ± 0.013 5 26.67 ± 0.10 3.342 ± 0.012 7 27.09 ± 0.103.291 ± 0.012 3 27.69 ± 0.10 3.221 ± 0.011 2 28.04 ± 0.10 3.183 ± 0.0113 28.93 ± 0.10 3.086 ± 0.010 5 29.84 ± 0.10 2.994 ± 0.010 2

Crystalline Form III, and all other crystalline forms of17α-ethynyl-5α-androstane-3α,17β-diol such as Form I, Form IV, Form V,Form VI, Form VII and Form VIII is optimally characterized at least inpart by reference to 1 or more, typically 2, 3, 4, 5, or 6 prominent,representative or characteristic peaks in the XRPD pattern by referenceto peak positions (degrees 2-theta) and optionally to peak intensities.

Differential scanning calorimetry (DSC) and TGA thermograms for Form IIIare presented in FIG. 3. TGA shows negligible weight loss from about 35°C. to about 105° C. indicating that the polymorphic forms within thesolid state mixture are most likely anhydrates. DSC shows a prominentendotherm at about 164° C. and is otherwise featureless, furtherindicating anhydrous material.

FIG. 4 is a proton FT-NMR (CDCl₃) of Form III17α-ethynyl-5α-androstane-3α,17β-diol, which does not exhibit additionalresonances due to solvent, thus further confirming that Form III is nota solvate.

FIG. 6 is a Raman spectroscopy spectrum of Form III17α-ethynyl-5α-androstane-3α,17β-diol Peak position for this Ramanspectrum is given in Table 2B.

TABLE 2B Position (cm⁻¹) Intensity 131.4 0.983 206.2 1.448 236.0 2.170272.3 0.808 324.1 0.736 373.9 0.806 384.0 0.858 395.4 1.003 432.1 1.040454.7 0.879 466.7 0.672 489.7 1.598 510.4 0.517 526.1 1.600 549.1 1.194602.5 1.328 619.2 1.152 648.3 1.225 657.5 0.978 702.9 1.360 733.2 2.078797.6 0.392 825.1 0.598 833.0 0.670 874.3 0.327 890.8 0.631 907.5 0.584928.8 0.459 963.3 0.825 986.2 0.430 1006.9 0.754 1038.0 0.615 1051.50.554 1093.1 0.362 1110.6 0.655 1150.0 1.014 1166.4 1.105 1189.8 0.7751235.8 0.972 1254.5 1.603 1293.4 0.592 1317.3 0.559 1335.1 0.863 1352.80.751 1376.3 0.686 1386.0 0.691 1434.8 1.856 1457.4 1.468 1469.5 0.9692103.9 2.626 2648.4 0.150 2857.1 1.486 2864.7 1.509 2942.9 2.821 2964.21.509 2982.8 0.774 3303.4 0.581

Calculated theoretical percentages of Compound 1 as the anhydrous,hemi-hydrate, and monohydrate forms compared with the measured elementalanalysis values are given in Table 3. The measured percent values forthe solid state obtained from the procedure described directly above lotare within the American Chemical Society recommendation values of 0.4%for the theoretical carbon and hydrogen percentage for anhydrousCompound 1. Therefore, elemental analysis confirms crystalline Form IIIobtained from the immediately proceeding procedure is an anhydrous solidstate form of Compound 1, as shown by the DSC/TGA thermograms and asfurther confirmed by low residual water determination of 0.20% by KarlFischer analysis.

TABLE 3 Calculated v Actual % C and H for Form III Mol. Form. C₂₁H₃₀O₃C₂₁H₃₀O₃*1/2H₂O C₂₁H₃₀O₃*H₂O Measured Carbon % 79.70 77.49 75.40 79.3Hydrogen 10.11 10.22 10.25 10.3 %

Since crystalline Form III 17α-ethynyl-androstane-3α,17β-diol is ananhydrate, it does not contain any organic solvent such as methanol ordioxane, and thus it has the advantage of being free of additionalorganic material that can affect the biological activity of Compound 1or that may increase the intrinsic toxicity of the compound. Thus, inanimal studies where relatively high levels of17α-ethynyl-androstane-3α,17β-diol can be used to characterizeanti-tumor activity or toxicity, e.g., about 60 mg/kg or about 100 mg/kgin mice or rats, such high levels can contribute significant amounts ofa solvent such as methanol or dioxane in the solvate. Such organicmolecules can affect the 17α-ethynyl-androstane-3α,17β-diol itself invivo when such solvents, e.g., induce or otherwise modulate liverenzymes that adversely affect 17α-ethynyl-androstane-3α,17β-diolmetabolism or disposition. However, anhydrates may have decreasedthermodynamic stability in relationship to a corresponding hydrate,which is exhibited by hygroscopicity, particularly at high relativehumidity. Hygroscopic materials may also be problematic due todecreasing potency in a drug substance due to its increasing weight aswater is absorbed or to instability of a drug product or unit dosageform of the drug product (e.g., tablet crumbling).

Example 3

Determination of Form III 17α-ethynyl-5α-androstane-3α,17β-diol UnitCell Parameters by XRPD Pattern Indexing: Indexing of the highresolution XRPD pattern by a indexing method described elsewhere in thespecification, provides the indexing solution given in Table 4.

TABLE 4 Indexing Solution for XRPD Pattern of Form III Compound 1Trigonal/Hexagonal Trigonal P3₁ 12 (#151)/P3₂ 12 (#153) Family and P3₁(#144)/P3₂ P3₁21 (#152)/P3₂21 (#154) Space Group (#145) P6₂ (#171)/P6₄(#172) Z′/Z 2/6 1/6 a (Å) 10.962 b (Å) 10.962 c (Å) 27.356 α (deg) 90 β(deg) 90 γ (deg) 120 Volume (Å³/cell) 2846.8 V/Z (Å³/asym. unit) 474.5Assumed Composition C₂₁H₃₂O₂ Density (g/cm³) 1.11

The crystal form for Form III 17α-ethynyl-5α-androstane-3α,17β-diol maybe further indicated by the photograph of FIG. 8 obtained frommicroscopic examination of a sample of Form III.

Example 4 Preparation and Analysis of Crystalline Form I17α-ethynyl-5α-androstane-3α,17β-diol (Form I Compound 1)

Form I 17α-ethynyl-5α-androstane-3α,17β-diol was prepared by dissolvingCompound 1 (32 mg) methanol (200 μL) at 65° C. using an oil bat. Thesolution was warm filtered through 0.2 μm nylon filter into a cleanvial, which was then replaced in the oil bath. Water (200 μL) was addedto the solution, which caused some precipitation of solids. To increaseyield, the sample was refrigerated to 5° C. The resulting solids werecollected by vacuum filtration. Form I may also be prepared from FormIII according to a procedure given in a subsequent example.

The high resolution XRPD for Form I prepared from Form III is given inFIG. 9. Listing of observed peaks for the XRPD pattern of Form I isprovided in Table 5. Prominent peaks are at 10.59, 12.33, 14.29, 14.72,16.04, 16.41, 17.49, 20.27, 24.04 and 27.21±0.10 degrees 2θ.

TABLE 5 Peak Listing for XRPD Pattern of Form I-High Resolution °2θ dspace (Å) Intensity (%)  6.92 ± 0.10 12.770 ± 0.187  1  8.73 ± 0.1010.133 ± 0.117  1  9.69 ± 0.10 9.130 ± 0.095 1 10.59 ± 0.10 8.354 ±0.079 21 11.34 ± 0.10 7.802 ± 0.069 1 12.33 ± 0.10 7.180 ± 0.058 1113.46 ± 0.10 6.576 ± 0.049 2 13.73 ± 0.10 6.449 ± 0.047 4 14.29 ± 0.106.197 ± 0.043 30 14.72 ± 0.10 6.019 ± 0.041 100 15.79 ± 0.10 5.614 ±0.036 5 15.91 ± 0.10 5.570 ± 0.035 11 16.04 ± 0.10 5.526 ± 0.034 4816.41 ± 0.10 5.401 ± 0.033 13 17.49 ± 0.10 5.070 ± 0.029 24 17.73 ± 0.105.004 ± 0.028 1 18.29 ± 0.10 4.852 ± 0.026 5 19.15 ± 0.10 4.634 ± 0.0246 19.46 ± 0.10 4.563 ± 0.023 1 19.98 ± 0.10 4.444 ± 0.022 4 20.27 ± 0.104.380 ± 0.021 13 21.15 ± 0.10 4.200 ± 0.020 1 22.42 ± 0.10 3.965 ± 0.0183 22.58 ± 0.10 3.938 ± 0.017 3 23.78 ± 0.10 3.743 ± 0.016 3 24.04 ± 0.103.702 ± 0.015 16 24.41 ± 0.10 3.647 ± 0.015 1 24.79 ± 0.10 3.591 ± 0.0141 25.15 ± 0.10 3.540 ± 0.014 8 25.41 ± 0.10 3.505 ± 0.014 6 25.77 ± 0.103.457 ± 0.013 1 26.10 ± 0.10 3.414 ± 0.013 4 26.62 ± 0.10 3.349 ± 0.0121 27.21 ± 0.10 3.277 ± 0.012 15 27.64 ± 0.10 3.227 ± 0.011 1 27.96 ±0.10 3.191 ± 0.011 1 28.43 ± 0.10 3.140 ± 0.011 1 28.81 ± 0.10 3.099 ±0.011 2 29.04 ± 0.10 3.075 ± 0.010 4 29.37 ± 0.10 3.042 ± 0.010 1 29.67± 0.10 3.011 ± 0.010 2

DSC and TGA thermograms for Form I 17α-ethynyl-5α-androstane-3α,17β-diolusing a temperature ramp of 10° C./min, is presented in FIG. 10. The TGAthermogram shows a weight loss of about 12% between about 60° C. andabout 105° C. that is accompanied by an endotherm at 88° C. in the DSCthermogram to form what may be a desolvate or partial desolvate of FormI. The DSC additionally shows a endotherm at 115° C. that is notassociated with TGA weight loss and may represent latticerearrangement(s) of a desolvated form with or without involvement of afurther underlying desolvation event. The DSC further exhibits aprominent endotherm at about 164° C.

The proton FT-NMR (CDCl₃) spectrum of Form I17α-ethynyl-5α-androstane-3α,17β-diol exhibits an additional resonance(δ 3.49 ppm) as compared the proton NMR spectrum of anhydrate Form III,which is due to methanol solvent. Thus NMR spectroscopy indicates thatForm I is at least a methanol solvate, which is supported by the DSC/TGAdata for Form I. FIG. 11 is a Raman spectroscopy spectrum of Form I17α-ethynyl-5α-androstane-3α,17β-diol. Peak position for this Ramanspectrum is given in Table 5B.

TABLE 5B Position (cm⁻¹) Intensity 147.8 1.523 231.3 2.136 324.5 0.628342.3 0.696 387.4 0.830 399.5 0.912 429.7 1.063 457.3 0.989 466.7 0.837493.6 0.761 528.5 1.191 548.5 0.745 600.6 1.100 613.9 1.081 647.8 1.070657.6 1.053 702.9 1.378 730.1 1.912 796.3 0.430 809.2 0.406 831.2 1.151890.3 0.628 904.7 0.382 916.2 0.442 929.5 0.581 965.5 0.711 1000.0 0.9061024.7 1.275 1076.1 0.422 1090.8 0.459 1107.0 0.663 1112.6 0.647 1148.21.310 1169.8 1.170 1188.3 0.631 1230.5 0.940 1253.8 1.500 1281.1 0.6781297.7 0.564 1314.1 0.451 1332.3 0.749 1342.5 0.791 1362.4 0.754 1383.70.567 1438.1 1.658 1455.5 1.799 2104.2 4.072 2856.5 2.259 2932.2 3.3782957.5 3.178 2975.8 1.537 3306.9 0.266

Indexing of the high resolution XRPD pattern by a indexing methoddescribed elsewhere in the specification and comparison to the indexingsolution of Form III, indicates that Form I is a methanol solvate withadditional presence of water, which remains consistent with the DSC/TGAdata obtained for Form I. Thus, Form I is a mixed methanol:water solvateof Compound 1. Karl-Fischer (KF) analysis indicates between about 5%water. Single crystal X-ray crystallography, described in a subsequentexample, on Form I 17α-ethynyl-5α-androstane-3α,17β-diol confirms Form Ito be a methanol-water solvate which has solvate stoichiometry of 1:1:1Compound 1:methanol:water.

Crystalline Form I 17α-ethynyl-androstane-3α,17β-diol has the advantageof being a relatively thermodynamically stable pseudopolymorph in thepresence of water. Typically, the most stable polymorphic form is chosenfor commercial development to circumvent problems from changes incrystalline form that may occur during preparation of the drug substanceor drug product or on storage of the drug product to ensure adequateshelf-life. However, less thermodynamically stable polymorphic forms maybe desirable due to their expected greater intrinsic dissolution rates,which could positively impact oral bioavailability. Additionally, if themost stable polymorphic form is a solvate (i.e. is a pseudopolymorph)there will be toxicology considerations if the solvate is other than ofwater. If the solvate is of water (i.e., the pseudopolymorph is ahydrate) other considerations come into play such as expected decreaseof intrinsic dissolution rate of hydrates in aqueous solution ascompared to the corresponding anhydrate, which could negatively impactoral bioavailability.

Example 5 Conversion of Crystalline Form III17α-ethynyl-5α-androstane-3α,17β-diol to Form I

Form III Compound 1 (77.1 mg) was added to a glass vial, followed byMeOH (0.8 mL) and a stirring bar. The sample was placed in an oil bathat 65° C. and solids completely dissolved. Additional Form III was addedso that excess solids remained and the mixture stirred for about 20minutes. Additional MeOH (0.2 mL) was added and the mixture was hotfiltered into a clean vial using a 0.2 μm nylon filter, and the filtratewas then placed in the oil bath at 65° C. Water (1 mL) was added and themixture cooled to 20° C. over 2 hours. XRPD analysis confirms that FormIII has converted to Form I as described by the immediately precedingexample. Form I was also produced from a slurry of Form III in 1:1methanol water or 3:1 methanol:water that is seeded with samples havingXRPD patterns of Form IV, Form V, Form VI, Form VII and Form VIII andstirred for 5 da at about 65° C. or 7 da at about 5° C. Thisinterconversion of Form III to Form I, in the presence of various othercrystalline forms with Form I excluded indicates that Form I is the moststable polymorphic form in methanol-water compared to Forms IIII-VIII.

Example 6 Single X-Ray Crystallography of Crystalline Form I17α-ethynyl-5α-androstane-3α,17β-diol (Form I Compound 1)

Single Crystal Preparation:

A single crystal of Form I suitable for single crystal X-raycrystalography was prepared by a slow cool of a solution of Compound 1in MeOH-water (1:1) to refrigerator temperature.

Data Collection:

A colorless needle of C₂₂H₃₈O₄ (C₂₁H₃₂O₂.1CH₃OH.1H₂O) having approximatedimensions of 0.25×0.08×0.06 mm, was mounted on a glass fiber in randomorientation. Preliminary examination and data collection were performedwith Cu K_(α) radiation (λ=1.54184 Å) on a Rigaku Rapid II™diffractometer equipped with a graphite crystal, incident beammonochromator. Refinements were performed on a LINUX™ PC using SHELX97™(Sheldrick, G. M. Acta Cryst., 2008, A64, 112). Cell constants and anorientation matrix for data collection were obtained from least-squaresrefinement using the setting angles of 2719 reflections in the range5°<θ<72°. The refined mosaicity from CRYSTALCLEAR™ (CrystalClear: “AnIntegrated Program for the Collection and Processing of Area DetectorData”, Rigaku Corporation, © 1997-2002) is 0.35° indicating good crystalquality. The space group was determined by the program XPREP™ (Bruker,XPREP in SHELXTL v. 6.12., Bruker AXS Inc., Madison, Wis., USA, 2002.From the systematic presence of the following conditions: h00 h=2n; 0k0k=2n; 00l l=2n, and from subsequent least-squares refinement, the spacegroup was determined to be P2₁2₁2₁ (No. 19). The data were collected toa maximum 2θ value of 143.69°, at a temperature of 298±1K.

Data Reduction:

Frames were integrated with CRYSTALCLEAR™. A total of 9981 reflectionswere collected, of which 3879 were unique. Lorentz and polarizationcorrections were applied to the data. The linear absorption coefficientis 0.577 mm⁻¹ for Mo K_(α) radiation. An empirical absorption correctionusing CRYSTALCLEAR™ was applied. Transmission coefficients ranged from0.856 to 0.966. Intensities of equivalent reflections were averaged. Theagreement factor for the averaging was 9.1% based on intensity.

Structure Solution and Refinement:

The structure was solved by direct methods using SIR2004 (Burla, M. C.,et al. J. Appl. Cryst. 2005, 38, 381).

The standard deviation of an observation of unit weight(goodness-of-fit) was 0.735. The highest peak in the final differenceFourier had a height of 0.16 e/Å³. The minimum negative peak had aheight of −0.17 e/Å³.

A calculated XRPD pattern was generated for Cu radiation usingPOWDERCELL™ 2.3 (Kraus, W.; Nolze, G. “PowderCell for Windows Version2.3” Federal Institute for Materials Research and Testing, BerlinGermany, EU, 1999) and compared to the experimentally acquired XRPD toconfirm the solution. FIG. 13 shows a comparison of the calculated XRPDpattern of Form I Compound 1 generated from single crystal data, withthe experimental pattern of Form I. All peaks in the experimentalpattern are represented in the calculated XRPD pattern, indicating thebulk material is likely a single phase. In general, differences inintensities between the calculated and experimental powder diffractionpatterns are typically due to preferred orientation. Preferredorientation is the tendency for crystals, usually plates or needles, tobe aligned with some degree of order. This preferred orientation of thesample can significantly change peak intensities, but not peakpositions, in the experimental powder diffraction pattern.

Single crystal data and data collection parameters are provided in Table6 and atomic coordinates are in Table 7. The orthorhombic cellparameters and calculated volume are: a=7.4893 (4) Å, b=11.0586 (8) Å,c=25.5095 (15) Å, α=90.00°, β=90.00°, γ=90.00°, V=2112.7 (2) Å³. Theformula weight of the asymmetric unit in the crystal structure of Form ICompound 1 is 366.55 amu·(formula unit)⁻¹ with Z=4, results in acalculated density of 1.152 g cm⁻³. The space group was determined to beP2₁2₁2₁. The quality of the structure obtained is high, as indicated bythe R-value of 0.049 (4.9%). Usually R-values in the range of 0.02 to0.06 are quoted for the most reliably determined structures Glusker, J,P., et al. “Crystal Structure Analysis: A Primer”, 2^(nd) ed.; OxfordUniversity press: New York, 1985; p. 87.

TABLE 6 Crystal Data and Data Collection Parameters for Form I Compound1 formula C₂₂H₃₈O₄ formula weight 366.55 space group P2₁2₁2₁ (No. 19) a,A 7.4893(4) b, Å 11.0586(8) c, Å 25.5095(15) V, Å³ 2112.7(2) Z 4d_(calc), g cm⁻³ 1.152 crystal dimensions, mm 0.25 × 0.08 × 0.06temperature, K 298. radiation (wavelength, Å) Cu K_(a) (1.54184)monochromator graphite linear abs coef, mm⁻¹ 0.577 absorption correctionapplied empirical^(a) transmission factors: min, max 0.856, 0.966diffractometer Nonius KappaCCD h, k, l range −9 to 6 −8 to 13 −28 to 312θ range, deg 10.40-143.69 mosaicity, deg 0.35 programs used SHELXTLF₀₀₀ 808.0 weighting 1/[s²(F_(o) ²) + (0.0151P)² + 0.0000P] where P =(F_(o) ² + 2F_(c) ²)/3 data collected 9981 unique data 3879 R_(int)0.091 data used in refinement 3879 cutoff used in R-factor calculationsF_(o) ² > 2.0 s(F_(o) ²) data with I > 2.0 s(I) 1501 number of variables258 largest shift/esd in final cycle 0.00 R(F_(o)) 0.049 R_(w)(F_(o) ²)0.077 goodness of fit 0.735

TABLE 7 Positional Parameters and Their Estimated Standard Deviationsfor Form I Compound 1 Atom x y z U(Å²) O1W −0.0438(4) 0.8199(3)0.96234(12) 0.0749(12) O31 −0.3061(4) 1.3642(3) 0.52670(11) 0.0586(9)O171 −0.2218(3) 1.8239(3) 0.86469(9) 0.0557(9) O911 0.4544(4) 1.7308(3)0.89214(13) 0.0793(10) C1 −0.4852(4) 1.5524(3) 0.59621(10) 0.0497(10) C2−0.5224(4) 1.5235(3) 0.53815(10) 0.0505(12) C3 −0.3548(5) 1.4846(3)0.50871(12) 0.0528(12) C4 −0.2024(4) 1.5732(3) 0.51687(11) 0.0474(9) C5−0.1678(4) 1.5973(3) 0.57550(11) 0.0411(9) C6 −0.0029(4) 1.6772(3)0.58395(10) 0.0489(10) C7 0.0429(4) 1.6841(3) 0.64227(10) 0.0452(10) C8−0.1187(4) 1.7286(3) 0.67539(10) 0.0387(9) C9 −0.2860(4) 1.6515(3)0.66401(10) 0.0372(9) C10 −0.3361(4) 1.6463(3) 0.60464(11) 0.0387(9) C11−0.4445(3) 1.6896(3) 0.69980(10) 0.0442(9) C12 −0.3944(4) 1.6880(3)0.75845(10) 0.0483(9) C13 −0.2289(4) 1.7654(3) 0.76893(11) 0.0379(9) C14−0.0767(4) 1.7226(3) 0.73391(11) 0.0414(9) C15 0.0892(4) 1.7883(3)0.75501(11) 0.0570(12) C16 0.0511(4) 1.8020(3) 0.81432(11) 0.0577(12)C17 −0.1400(4) 1.7528(3) 0.82396(12) 0.0459(10) C18 −0.2752(4) 1.8999(3)0.76101(11) 0.0570(10) C19 −0.3946(4) 1.7704(3) 0.58417(10) 0.0535(10)C171 −0.1343(6) 1.6254(4) 0.84154(12) 0.0592(13) C172 −0.1224(7)1.5266(4) 0.85662(14) 0.0917(17) C912 0.4047(6) 1.6084(4) 0.89996(14)0.0987(18) H31 −0.219(7) 1.347(5) 0.521(2) 0.18(3)* H171 −0.311(5)1.785(4) 0.8736(14) 0.105(18)* H1W1 −0.081(5) 0.767(3) 0.9783(13)0.072(15)* H1W2 −0.082(5) 0.835(4) 0.9320(13) 0.119(18)* H911 0.441(7)1.770(4) 0.9170(16) 0.14(2)* H3 −0.382 1.481 0.471 0.064 H5 −0.140 1.5190.591 0.049 H8 −0.144 1.813 0.666 0.046 H9 −0.255 1.569 0.674 0.045 H14−0.059 1.637 0.742 0.050 H1A −0.594 1.582 0.612 0.060 H1B −0.452 1.4780.614 0.060 H2A −0.610 1.459 0.536 0.061 H2B −0.572 1.595 0.521 0.061H4A −0.231 1.649 0.499 0.057 H4B −0.095 1.541 0.501 0.057 H6A 0.0971.644 0.565 0.059 H6B −0.026 1.758 0.571 0.059 H7A 0.142 1.739 0.6470.054 H7B 0.079 1.605 0.655 0.054 H11A −0.483 1.770 0.690 0.053 H11B−0.544 1.635 0.694 0.053 H12A −0.494 1.718 0.779 0.058 H12B −0.371 1.6050.769 0.058 H15A 0.103 1.867 0.738 0.069 H15B 0.196 1.741 0.749 0.069H16A 0.058 1.886 0.825 0.069 H16B 0.137 1.756 0.835 0.069 H172 −0.1131.447 0.869 0.110 H18A −0.369 1.922 0.785 0.085 H18B −0.314 1.913 0.7250.085 H18C −0.171 1.948 0.768 0.085 H19A −0.506 1.793 0.600 0.080 H19B−0.410 1.767 0.547 0.080 H19C −0.305 1.829 0.593 0.080 H91A 0.507 1.5570.896 0.148 H91B 0.315 1.586 0.875 0.148 H91C 0.358 1.599 0.935 0.148Starred atoms were refined isotropically U_(eq) = (⅓)Σ_(i)Σ_(j)U_(ij)a*_(i)a*_(j)a_(i) · a_(j) Hydrogen atoms are included incalculation of structure factors but not refined

ORTEP representation of the Form I unit cell is given by FIG. 14. TheORTEP diagram was prepared using ORTEP III™ program (Johnson, C. K.ORTEP III, Report ORNL-6895, Oak Ridge National Laboratory, TN, U.S.A.1996; Farrugia, L. J. “OPTEP-3 for Windows V1.05”, J. Appl. Cryst. 1997,30, 565) within the PLATON software package (Spek, A. L. “PLATON.Molecular Graphics Program” Utrecht University, Utrecht, TheNetherlands, 2008; Spek, A. L, J. Appl. Cryst. 2003, 36, 7). Atoms arerepresented by 50% probability anisotropic thermal ellipsoids. Packingdiagrams were prepared using CAMERON™ modeling software (Watkin, D. J.;Prout, C. K.; Pearce, L. J. CAMERON, Chemical CrystallographyLaboratory, University of Oxford, Oxford, 1996).

The molecule observed in the asymmetric unit of the single crystalstructure is consistent with the molecular structure of Compound 1,although FIG. 14 displays the opposite stereochemical configuration.This configuration was arbitrarily chosen, since the absoluteconfiguration could not be assigned as the crystal was a racemic twin.The asymmetric unit shown in FIG. 14 contains one molecule of Compound 1with one water molecule and one methanol molecule.

Example 7 Preparation and Analysis of Crystalline Form IV17α-ethynyl-androstane-3α,17β-diol (Form IV Compound 1)

Solids with XRPD pattern of Form IV was prepared from fast evaporationof a solution of Compound 1 in 2:3 acetonitile:water solution or fromcrash precipitation of an acetonitrile solution of Compound 1 by addingwater. Alternatively, Form IV was prepared form Compound 1 (34 mg)dissolved in acetonitrile (6 mL). This solution was filtered through 0.2μm nylon filter into a clean vial, whereupon water (9 mL) was added. Theresulting solution was allowed to evaporate under ambient conditions andthe solids were isolated by vacuum filtration. Form IV was also preparedby dissolving Compound 1 (31 mg) into acetone (500 μL), filtering thesolution through 0.2 μm nylon filter into a clean vial, and then addingwater (250 μL). The solution so formed is allowed to evaporate underambient conditions. Solids were isolated by vacuum filtration.

Needle crystal morphology was found for well formed crystals under 10×magnification with bifringence and extinction observed under polarizedlight due the anisotropic nature of these crystals.

The low resolution XRPD for Form IV prepared by the immediatelypreceding procedure is given in FIG. 15. Listing of observed peaks forthe XRPD pattern of Form IV is provided in Table 8. Prominent peaks areat 8.31, 9.84, 11.28, 13.02, 13.86, 14.73, 15.00, 16.14, 16.53, 17.01,17.76 and 18.72±0.10 degrees 2θ.

TABLE 8 Peak Listing for XRPD Pattern of Form IV-Low Resolution °2θ dspace (Å) Intensity (%)  8.31 ± 0.10 10.640 ± 0.129  35  8.91 ± 0.109.925 ± 0.112 5  9.84 ± 0.10 8.989 ± 0.092 82 11.28 ± 0.10 7.844 ± 0.07039 12.36 ± 0.10 7.161 ± 0.058 10 13.02 ± 0.10 6.800 ± 0.052 57 13.86 ±0.10 6.390 ± 0.046 28 14.25 ± 0.10 6.216 ± 0.044 11 14.73 ± 0.10 6.014 ±0.041 93 15.00 ± 0.10 5.906 ± 0.039 66 15.36 ± 0.10 5.769 ± 0.038 2216.14 ± 0.10 5.492 ± 0.034 70 16.53 ± 0.10 5.363 ± 0.032 75 17.01 ± 0.105.213 ± 0.031 100 17.76 ± 0.10 4.994 ± 0.028 35 18.12 ± 0.10 4.896 ±0.027 11 18.72 ± 0.10 4.740 ± 0.025 34 19.35 ± 0.10 4.587 ± 0.024 420.04 ± 0.10 4.431 ± 0.022 21 20.31 ± 0.10 4.373 ± 0.021 6 20.76 ± 0.104.279 ± 0.020 15 21.36 ± 0.10 4.160 ± 0.019 2 22.14 ± 0.10 4.015 ± 0.01820 22.95 ± 0.10 3.875 ± 0.017 3 23.67 ± 0.10 3.759 ± 0.016 8 23.79 ±0.10 3.740 ± 0.016 7 24.27 ± 0.10 3.667 ± 0.015 3 24.63 ± 0.10 3.615 ±0.015 6 25.14 ± 0.10 3.542 ± 0.014 18 26.10 ± 0.10 3.414 ± 0.013 6 26.73± 0.10 3.335 ± 0.012 7 27.24 ± 0.10 3.274 ± 0.012 7 27.66 ± 0.10 3.225 ±0.011 12 28.41 ± 0.10 3.142 ± 0.011 8 28.92 ± 0.10 3.087 ± 0.010 7

DTA and TGA thermograms for Form IV17α-ethynyl-5α-androstane-3α,17β-diol using a temperature ramp of 10°C./min, are presented in FIG. 16. The TGA thermogram for Form IV showsbetween about 5-6% weight loss from about 60° C. to about 105° C. orabout 7% weight loss from between about 40° C. to about 160° C.concomitant with a broad endotherm in the DTA thermogram centered atabout 88° C. followed by an endotherm at about 106° C. These transitionsare consistent with a monohydrate pseudopolymorph than undergoestransition to a more thermodynamically stable polymorphic form followingits dehydration. A prominent endotherm is then observed at about 164° C.

Since crystalline Form IV 17α-ethynyl-androstane-3α,17β-diol is amonohydrate, it does not contain an organic solvent such as methanol ordioxane and thus has the advantage of being free of organic materialthat can affect biological activity of Compound 1 or that may increaseits intrinsic toxicity. The presence of water in the crystal lattice isexpected to contribute to this crystal form's stability with respect toabsorbing water on prolonged storage because water is already present inthe crystal lattice and thus should not be as hygroscopic as thecorresponding anhydrate or a lower hydrate. Based on the expectedphysical property of a solid with the morphology of FIG. 17, crystallineForm IV is also expected to have a favorable intrinsic dissolution ratedue its high surface to volume ratio compared to other crystalmorphologies. Flow characteristics for mechanical manipulations (e.g.,blending), however, may not be as favorable compared to other crystalmorphologies due to the directional asymmetry of the crystal shape.

Example 8 Preparation of Crystalline Form V17α-ethynyl-androstane-3α,17β-diol (Form V Compound 1)

Solids having the XRPD pattern of Form V was obtained by adding Form IIICompound 1 (41 mg) to 2,2,2-trifluoroethanol (1600 μL) Additional FormIII Compound 1 was added to form a slurry that was stirred at 30° C. for6 days. Solids were isolated by vacuum filtration, air-dried andanalyzed by XRPD. No definite crystal morphology was found for thesolids so formed at 10× magnification and bifringence and extinctionunder polarized light was not observed, which indicates a highlydisordered crystalline state for this solid state form.

The low resolution XRPD for Form V prepared by the immediately precedingprocedure is given in FIG. 18. Listing of observed peaks for the XRPDpattern of Form V is provided in Table 9. Prominent peaks are at 5.82,9.48, 11.49, 13.50, 15.21, 17.28 and 18.93±0.10 degrees 2θ.

TABLE 9 Peak Listing for XRPD Pattern of Form V-Low Resolution °2θ dspace (Å) Intensity (%)  5.82 ± 0.10 15.186 ± 0.265  81  8.04 ± 0.1010.997 ± 0.138  9  9.48 ± 0.10 9.330 ± 0.099 90 11.49 ± 0.10 7.702 ±0.067 80 12.24 ± 0.10 7.231 ± 0.059 2 13.50 ± 0.10 6.559 ± 0.049 3814.04 ± 0.10 6.308 ± 0.045 7 14.34 ± 0.10 6.177 ± 0.043 5 15.21 ± 0.105.825 ± 0.038 78 15.99 ± 0.10 5.543 ± 0.035 23 16.47 ± 0.10 5.382 ±0.033 23 17.28 ± 0.10 5.132 ± 0.030 65 18.93 ± 0.10 4.688 ± 0.025 10019.32 ± 0.10 4.594 ± 0.024 18 20.85 ± 0.10 4.261 ± 0.020 27 21.66 ± 0.104.103 ± 0.019 6 22.41 ± 0.10 3.967 ± 0.018 5 23.01 ± 0.10 3.865 ± 0.01713 23.22 ± 0.10 3.831 ± 0.016 5 23.61 ± 0.10 3.768 ± 0.016 3 24.12 ±0.10 3.690 ± 0.015 6 24.51 ± 0.10 3.632 ± 0.015 27 25.32 ± 0.10 3.518 ±0.014 7 25.65 ± 0.10 3.473 ± 0.013 4 26.13 ± 0.10 3.410 ± 0.013 4 26.61± 0.10 3.350 ± 0.012 2 27.33 ± 0.10 3.263 ± 0.012 16 27.75 ± 0.10 3.215± 0.011 5 28.05 ± 0.10 3.181 ± 0.011 5 29.25 ± 0.10 3.053 ± 0.010 21

TGA thermogram for Form V 17α-ethynyl-5α-androstane-3α,17β-diol using atemperature ramp of 10° C./min shows negligible weight loss from about40° C. to about 160° C. while the DTA thermogram shows a prominentendotherm at about 164° C. and is otherwise featureless. Thus,crystalline Form V of 17α-ethynyl-androstane-3α,17β-diol is an anhydrate(i.e., has no solvent in its crystal structure). On prolonged standing,a sample of Form V was found to have undergone a polymorphic transitionto Form III as evidenced by XRPD reanalysis.

Compared to a solvate containing an organic solvent, this material isexpected to have the advantage of containing no organic solvent such asdioxane or methanol. In animal studies where relatively high levels of17α-ethynyl-androstane-3α,17β-diol may be used to characterizeanti-tumor or other biological activity or toxicity of the material,such high levels can contribute significant amounts of a solvent such asmethanol or dioxane in the solvate. Such organic molecules can affectthe 17α-ethynyl-androstane-3α,17β-diol itself in vivo when suchsolvents, e.g., induce or otherwise modulate liver enzymes thatadversely affect 17α-ethynyl-androstane-3α,17β-diol or disposition.However, anhydrates may have decreased thermodynamic stability inrelationship to a corresponding hydrate, which is exhibited byhygroscopicity, particularly at high relative humidity. Hygroscopicmaterials may also be problematic due to decreasing potency in a drugsubstance due to its increasing weight as water is absorbed or toinstability of a drug product or unit dosage form of the drug product(e.g., tablet crumbling). Offsetting such disadvantages is thedisordered crystalline state of Form V that may prove advantageous dueto an expected higher intrinsic dissolution rate, since there will beless crystalline lattice forces to be overcome, compared to acrystalline form with highly ordered crystalline state.

Example 9 Preparation and Analysis of Crystalline Form VI17α-ethynyl-androstane-3α,17β-diol (Form VI Compound 1)

Form VI was prepared by dissolving Compound 1 (66 mg) in dioxane (2800μL) with sonication. The solution was filtered through 0.2 μm nylonfilter into a clean vial and allowed to evaporate under ambientconditions, yielding a gel, which was then dried under vacuum for 1 dayat ambient temperature.

Irregular crystal fragments were observed, but no definite crystalmorphology could be discerned at 10× magnification; however, bifringenceand extinction under polarized light was observed, which indicates ananisotropic crystalline shape that may appear highly disordered in bulkamount due to the presence of significant numbers of crystal defects.

The low resolution XRPD for Form VI prepared by the immediatelypreceding procedure is given in FIG. 19. Listing of observed peaks forthe XRPD pattern of Form VI is provided in Table 10. Prominent peaks are7.17, 9.78, 13.26, 14.25, 14.61, 15.000 and 18.69±0.10 degrees 2θ.

TABLE 10 Peak Listing for XRPD Pattern of Form VI-Low Resolution °2θ dspace (Å) Intensity (%)  3.60 ± 0.10 24.544 ± 0.701  6  6.69 ± 0.1013.213 ± 0.200  16  7.17 ± 0.10 12.329 ± 0.174  36  7.50 ± 0.10 11.787 ±0.159  19  9.15 ± 0.10 9.665 ± 0.107 4  9.78 ± 0.10 9.044 ± 0.093 4313.26 ± 0.10 6.677 ± 0.051 100 13.95 ± 0.10 6.348 ± 0.046 21 14.25 ±0.10 6.216 ± 0.044 57 14.61 ± 0.10 6.063 ± 0.042 69 15.00 ± 0.10 5.906 ±0.039 44 15.39 ± 0.10 5.758 ± 0.037 9 15.99 ± 0.10 5.543 ± 0.035 1416.44 ± 0.10 5.392 ± 0.033 9 16.83 ± 0.10 5.268 ± 0.031 8 17.04 ± 0.105.204 ± 0.030 17 17.28 ± 0.10 5.132 ± 0.030 11 17.67 ± 0.10 5.019 ±0.028 16 18.27 ± 0.10 4.856 ± 0.026 25 18.69 ± 0.10 4.748 ± 0.025 8419.53 ± 0.10 4.545 ± 0.023 14 19.92 ± 0.10 4.457 ± 0.022 6 20.31 ± 0.104.373 ± 0.021 7 20.52 ± 0.10 4.328 ± 0.021 9 20.91 ± 0.10 4.248 ± 0.02018 21.39 ± 0.10 4.154 ± 0.019 12 21.57 ± 0.10 4.120 ± 0.019 12 21.78 ±0.10 4.081 ± 0.019 20 22.65 ± 0.10 3.926 ± 0.017 11 23.55 ± 0.10 3.778 ±0.016 8 23.91 ± 0.10 3.722 ± 0.015 11 24.66 ± 0.10 3.610 ± 0.014 7 25.50± 0.10 3.493 ± 0.014 7 26.01 ± 0.10 3.426 ± 0.013 3 26.28 ± 0.10 3.391 ±0.013 3 26.61 ± 0.10 3.350 ± 0.012 3 27.12 ± 0.10 3.288 ± 0.012 5 27.39± 0.10 3.256 ± 0.012 6 28.17 ± 0.10 3.168 ± 0.011 7 29.10 ± 0.10 3.069 ±0.010 4 29.64 ± 0.10 3.014 ± 0.010 8

Thermal analysis of Form VI 17α-ethynyl-5α-androstane-3α,17β-diol shownin FIG. 20 provides a TGA thermogram with about 5% weight loss fromabout 40° C. to about 85° C. and about 12% weight loss from about 40° C.to about 180° C. and a DTA thermogram (after calibration correction)with a broad endotherm centered at about 70° C. immediately followed bya poorly defined broad exotherm. This thermal data is consistent with asolvate that desolvates with rearrangement of the crystal lattice toform a thermodynamically more stable desolvate or partial desolvate. TheDTA further shows a subsequent prominent endotherm at 164° C.

The ¹H-NMR (CDCl₃) spectrum of Form VI17α-ethynyl-5α-androstane-3α,17β-diol exhibits additional resonance (δ3.64 ppm) as compared the proton NMR spectrum of anhydrate Form III, dueto dioxane solvent. Prior to dissolution of Form VI for NMR analysis,the Form VI sample was washed with CCl₄ to remove non-specificallyabsorbed surface solvent. Proton NMR spectroscopy thus indicates Form VIis a dioxane solvate, which is supported by the DTA/TGA data, and ismost likely a mono dioxane solvate based upon integration of the δ 3.64ppm resonance and the weight loss in TGA. After prolonged standing atambient temperature and pressure, the Form VI sample was reanalyzed byDTA/TGA and proton NMR. The thermal reanalysis now shows no TGA weightloss and no desolvation events in DTA. Furthermore, proton NMR now showssignificant diminution of the δ 3.64 ppm resonance. Thus, desolvation ofForm VI may occur if storage precautions are inadequate and may beaccompanied by a polymorphic transition to a more thermodynamicallystable form such as Form III.

Form VI as a dioxane solvate is expected to be useful as an internalstandard for quantifying amounts of Compound 1 by proton-NMR or ¹³C-NMRspectroscopy in samples with an unknown content of Compound 1.Usefulness of Form VI as an internal standard in proton NMRspectroscopic analysis will be due to the presence of a singlet ofrelatively high intensity in the proton NMR spectrum due to eightmagnetically equivalent protons contributed by the dioxane solvent thatis in a precise 1:1 ratio to Compound 1 with respect to Form VI that isadded to the sample to be quantified. Usefulness of Form VI as aninternal standard in ¹³C NMR spectroscopic analysis will be due to thepresence of a singlet of relatively high intensity in a proton decoupled¹³C-NMR spectra due to four magnetically equivalent carbons contributedby the dioxane solvent that is in a precise 1:1 ratio to Compound 1 withrespect to Form VI that is added to the sample to be quantified.

Example 10 Preparation and Analysis of Crystalline Form VII17α-ethynyl-androstane-3α,17β-diol (Form VII Compound 1)

Form VII Compound 1 was prepared by dissolving Compound 1 in ethanol(1.2 mL) at 47° C. The solution was evaporated to half volume undernitrogen causing a solid mass to form. Isopropyl acetate (1 mL) was thenadded and the solids re-dissolved with sonication. The solution wasevaporated to half volume to form solids. The mixture was heated to 47°C., which caused complete dissolution and cooled to ambient temperature,causing a small amount of solids to form. The mixture so formed wasrefrigerated for several hours and the resulting solids were isolated byvacuum filtration.

Irregular agglomerated crystals were observed at 10× magnification thatdid not permit description of crystal morphology. Examination underpolarized light did exhibit domains having bifringence and extinction,which indicates an anisotropic crystalline shape that may appear highlydisordered in bulk amount due to the presence of significant numbers ofcrystal defects.

The low resolution XRPD for Form VII prepared from the immediatelypreceding procedure is given in FIG. 21. Listing of observed peaks forthe XRPD pattern of Form VII is provided in Table 11. Prominent peaksare at 5.91, 9.78, 13.47, 14.16, 15.78, 17.85, 19.50 and 21.45±0.10degrees 2θ.

TABLE 11 Peak Listing for XRPD Pattern of Form VII-Low Resolution °2θ dspace (Å) Intensity (%)  5.91 ± 0.10 14.955 ± 0.257  26  8.31 ± 0.1010.640 ± 0.129  19  9.78 ± 0.10 9.044 ± 0.093 100 10.41 ± 0.10 8.498 ±0.082 4 11.37 ± 0.10 7.783 ± 0.069 17 11.88 ± 0.10 7.450 ± 0.063 1912.24 ± 0.10 7.231 ± 0.059 4 13.47 ± 0.10 6.574 ± 0.049 44 14.16 ± 0.106.255 ± 0.044 49 14.46 ± 0.10 6.126 ± 0.042 18 15.54 ± 0.10 5.702 ±0.037 26 15.78 ± 0.10 5.616 ± 0.036 51 16.50 ± 0.10 5.373 ± 0.033 2216.71 ± 0.10 5.306 ± 0.032 16 16.98 ± 0.10 5.222 ± 0.031 5 17.43 ± 0.105.088 ± 0.029 7 17.85 ± 0.10 4.969 ± 0.028 31 18.69 ± 0.10 4.748 ± 0.0256 18.99 ± 0.10 4.673 ± 0.025 9 19.50 ± 0.10 4.552 ± 0.023 67 19.71 ±0.10 4.504 ± 0.023 21 20.52 ± 0.10 4.328 ± 0.021 13 21.45 ± 0.10 4.143 ±0.019 31 22.86 ± 0.10 3.890 ± 0.017 12 23.49 ± 0.10 3.787 ± 0.016 1024.03 ± 0.10 3.703 ± 0.015 12 24.63 ± 0.10 3.615 ± 0.015 7 25.02 ± 0.103.559 ± 0.014 8 25.26 ± 0.10 3.526 ± 0.014 7 25.68 ± 0.10 3.469 ± 0.0132 26.43 ± 0.10 3.372 ± 0.013 13 26.64 ± 0.10 3.346 ± 0.012 11 26.91 ±0.10 3.313 ± 0.012 23 27.75 ± 0.10 3.215 ± 0.011 8 28.08 ± 0.10 3.178 ±0.011 3 28.71 ± 0.10 3.110 ± 0.011 7

TGA thermogram for Form VII 17α-ethynyl-5α-androstane-3α,17β-diol usinga temperature ramp of 10° C./min shows between about 2% to negligibleweight loss from about 40° C. to about 160° C. while the DTA thermogramshows a prominent endotherm at about 164° C. and is otherwisefeatureless. Thus, crystalline Form VII of17α-ethynyl-androstane-3α,17β-diol is most likely an anhydrate (i.e.,has no solvent in its crystal structure). On prolonged standing, asample of Form VII was found to have undergone a polymorphic transitionto Form III as evidenced by XRPD reanalysis.

Compared to a solvate containing an organic solvent, this material isexpected to have the advantage of containing no organic solvent such asdioxane or methanol. In animal studies where relatively high levels of17α-ethynyl-androstane-3α,17β-diol may be used to characterizeanti-tumor or other biological activity or toxicity of the material,such high levels can contribute significant amounts of a solvent such asmethanol or dioxane in the solvate. Such organic molecules can affectthe 17α-ethynyl-androstane-3α,17β-diol itself in vivo when suchsolvents, e.g., induce or otherwise modulate liver enzymes thatadversely affect 17α-ethynyl-androstane-3α,17β-diol or disposition.However, anhydrates may have decreased thermodynamic stability inrelationship to a corresponding hydrate, which is exhibited byhygroscopicity, particularly at high relative humidity. Hygroscopicmaterials may also be problematic due to decreasing potency in a drugsubstance due to its increasing weight as water is absorbed or toinstability of a drug product or unit dosage form of the drug product(e.g., tablet crumbling).

Example 11 Preparation and Analysis of Crystalline Form VIII17α-ethynyl-androstane-3α,17β-diol (Form VIII Compound 1)

Form VIII was prepared from slow evaporation of a dichloromethanesolution of Compound 1. Visual examination under 10× magnification showsirregular shaped crystal fragments. The low resolution XRPD for FormVIII prepared by the immediately preceding procedure is given in FIG.22. Listing of observed peaks for the XRPD pattern of Form VIII isprovided in Table 12. Prominent peaks are at 11.13, 15.96, 16.62, 17.76and 18.75±0.10 degrees 2θ.

TABLE 12 Peak Listing for XRPD Pattern of Form VIII-Low Resolution °2θ dspace (Å) Intensity (%)  9.39 ± 0.10 9.419 ± 0.101 10 10.11 ± 0.10 8.750± 0.087 11 11.13 ± 0.10 7.950 ± 0.072 35 13.50 ± 0.10 6.559 ± 0.049 614.76 ± 0.10 6.002 ± 0.041 9 15.96 ± 0.10 5.553 ± 0.035 100 16.62 ± 0.105.334 ± 0.032 26 17.76 ± 0.10 4.994 ± 0.028 31 18.75 ± 0.10 4.733 ±0.025 28 19.08 ± 0.10 4.652 ± 0.024 14 20.16 ± 0.10 4.405 ± 0.022 522.29 ± 0.10 3.988 ± 0.018 10 22.86 ± 0.10 3.890 ± 0.017 11 23.73 ± 0.103.750 ± 0.016 22 24.00 ± 0.10 3.708 ± 0.015 10 24.30 ± 0.10 3.663 ±0.015 22 25.17 ± 0.10 3.538 ± 0.014 7 25.38 ± 0.10 3.509 ± 0.014 9 25.74± 0.10 3.461 ± 0.013 8 26.16 ± 0.10 3.407 ± 0.013 12 27.96 ± 0.10 3.191± 0.011 10 28.41 ± 0.10 3.142 ± 0.011 22 29.85 ± 0.10 2.993 ± 0.010 6

TGA thermogram for Form VIII 17α-ethynyl-5α-androstane-3α,17β-diol usinga temperature ramp of 10° C./min shows negligible weight loss from about40° C. to about 160° C. while the DTA thermogram shows a prominentendotherm at about 164° C. and is otherwise featureless. Thus,crystalline Form VIII of 17α-ethynyl-androstane-3α,17β-diol is mostlikely an anhydrate (i.e., has no solvent in its crystal structure).

Compared to a solvate containing an organic solvent, this material isexpected to have the advantage of containing no organic solvent such asdioxane or methanol. In animal studies where relatively high levels of17α-ethynyl-androstane-3α,17β-diol may be used to characterizeanti-tumor or other biological activity or toxicity of the material,such high levels can contribute significant amounts of a solvent such asmethanol or dioxane in the solvate. Such organic molecules can affectthe 17α-ethynyl-androstane-3α,17β-diol itself in vivo when suchsolvents, e.g., induce or otherwise modulate liver enzymes thatadversely affect 17α-ethynyl-androstane-3α,17β-diol or disposition.However, anhydrates may have decreased thermodynamic stability inrelationship to a corresponding hydrate, which is exhibited byhygroscopicity, particularly at high relative humidity. Hygroscopicmaterials may also be problematic due to decreasing potency in a drugsubstance due to its increasing weight as water is absorbed or toinstability of a drug product or unit dosage form of the drug product(e.g., tablet crumbling). Due to its disordered crystalline state FormVII may be expected to have a greater intrinsic dissolution ratecompared to more ordered crystalline forms.

Example 12 Preparation and Analysis of Amorphous17-ethynyl-5α-androstane-3α,17β-diol (Amorphous Compound 1)

Compound 1 (52.0 mg) was dissolved in 200 μL of methanol by heating inan 80° C. water bath. The solution was allowed to reach room temperaturewhereupon 20 μL of water was added with swirling. The solid so formedwas filtered, washed with cold methanol and dried under vacuum to give24.3 mg of the title material.

The XRPD pattern given in FIG. 23 is a halo with no distinctive peaks,which is indicative of amorphous material. The DTA/TGA thermogramsprovided in FIG. 24, show an endothermic event at about 81° C. in theDTA trace with about 7% decrease of weight in the TGA thermogram fromabout 35° C. to about 60° C., using a temperature ramp of 10 C/min.These thermal events are indicative of desolvation. An exothermic eventin the DTA trace is subsequently observed at about 120° C., whichindicates a transition from amorphous to a thermodynamically stablepolymorph has occurred. This polymorph, presumed to be Form III, thenexhibits a prominent endotherm at about 163° C.

Amorphous compound 1, with varying degrees of crystallinity was alsoobtained form melt-quenching Form III according to the followingprocedure. Form III was lightly crushed with a spatula in a vial thatwas then placed in an oilbath set at 175° C. A nitrogen stream was runthrough the vial and the vial kept in the oilbath until the sample hadmelted. The vial was then removed and quickly placed in a bathcontaining dry ice with acetone. The sample was placed in a freezerunder desiccant and analyzed by XRPD for crystallinity.

The propensity of amorphous material to convert to various crystallineforms was studied by vapor stressing melt-quenched Compound 1, preparedfrom Form III by the immediately preceding procedure. Studies at highhumidity indicate that amorphous Compound 1 will convert back to FormIII further indicating that Form III is the most thermodynamicallystable polymorphic form provided Form III is not exposed tomethanol-water.

Example 13 Preparation of Formulations Comprising17α-ethynyl-5α-androstane-3α,17β-diol in Solid State Form

The following Table are lists of ingredients used in preparation ofcapsule formulations containing Compound 1 in solid state form.

TABLE 13 Formulation Containing 25 mg Solid State Compound 1 % w/wmg/capsule Drug Substance Compound 1 micronized 13.9 25.0 ExcipientsSodium lauryl sulfate, NF 10.0 18.0 Microcrystalline cellulose, NF 65.6118 (Avicel PH 102) Crospovidone, NF (Polypasdone 10.0 18.0 XL-10)Magnesium stearate, NF 0.5 1.0 Total 100 180 Hard gelatin capsule # 2

TABLE 14 Formulation Containing 5 mg Compound 1 in Solid State Form %w/w mg/capsule Drug Substance Compound 1 micronized 2.8 5.0 ExcipientsSodium lauryl sulfate, NF 10.0 18.0 Microcrystalline cellulose, NF 76.7138 (Avicel PH 102) Crospovidone, NF (Polypasdone 10.0 18.0 XL-10)Magnesium stearate, NF 0.5 1.0 Total 100 180 Hard gelatin capsule # 2

The following are ingredient lists used in preparation of a suspensionformulation of Compound 1 in solid state form.

TABLE 15 Suspension Formulation Containing Compound 1 in Solid StateForm % w/w Drug Substance Compound 1 micronized 2-10 ExcipientsPolysorbate 80 2.0 Carboxymethycellulose (CMC) 0.1 Sodium Chloride 0.9Phenol 0.05 Deionized water Remainder

In the formulations above and in the following examples solid stateforms of Compound 1 (e.g. amorphous or crystalline Form III) arepreferably micronized to a mean volume weighted particle size (Dv, 50)of between about 3 to about 100 microns prior to blending withexcipients. In one embodiment, Polymorph Form III is micronized to givea particle size with (Dv, 90)=10 μm (particle size that contains 90%(volume weighted) of all the particles). Selection of appropriateparticle size is a tradeoff between improved bioavailability for a solidstate form of Compound 1 in a given formulation due to improveddissolution rate of solid state Compound 1 and increased manufacturingcost of the formulation as particle size decreases. For example,particle sizes with a mean volume weighted particle size or averagediameter of less than about 3 microns typically requires fluid bedmicronization (for example, see Julia Z. H, et al. “Fluid bedgranulation of a poorly water soluble, low density, micronized drug:comparison with high shear granulation” Int. J. Pharm. 237 (1-2): 1-14(2002), which is more costly than jet milling to a larger particle sizeand is a process more difficult to scale up.

With dosage strengths of less than 5 mg (e.g., 1 mg) pre-blending ofmicronized Compound 1 with a surface active agent such as sodium laurylsulfate is sometimes conducted prior to blending with the remainingexcipients in order to obtain a uniform distribution of Compound 1within the formulation.

Example 14 Prostate Cancer

Treatment of Androstenedione (AED)-stimulated CaP in a Tumor Model usinga Formulation Prepared from Crystalline Form III.

Castrated SCID mice (six week-old) were implanted with a 5 mg AED,60-day time-release pellet (Innovative Labs, Sarasota Fla.). After threedays, all mice were injected subcutaneously in the right flank with 100μL of 7.5×10⁶ LNCaP tumor cells in phenol red-free RPMI mixed 1:1 withMatrigel. Tumor volumes were measured weekly and calculated as a2×b/2with a being the width and b the length of the tumor in millimeters(reported as mm³).

To test the effect of dose on tumor incidence, a total of 48 castratedmale SCID mice were implanted with LNCaP tumor cells as described, andthe mice were randomly divided into 4 groups of 12 animals each toprovide vehicle control (30% cyclodextrin-sulfobutylether in water)group and treatment groups using 4 mg/mouse/day, 1 mg/mouse/day, and 0.4mg/mouse/day). Treatment groups used a liquid formulation prepared bydissolving a solid state form of 17α-ethynyl-androst-5-ene-3α,17β-diol(Compound 1) in vehicle, which provided 20 mg Compound 1 per mL in 30%cyclodextrin-sulfobutylether in water. The liquid formulation soprepared was administered 24 hours after tumor inoculation as a 200 μLintraperitoneal (ip) injection. All animals were dosed daily for 28consecutive days and tumor volumes were measured weekly.

The results of this study (FIG. 25) showed a significant reduction intumor incidence (compared to vehicle) in the two highest dose groups, (1mg, p=0.006, n=11; 4 mg, p<0.001, n=12), with decreases in tumor volumeapparent in all three dose groups. There was a statistically significantdelay in the time to a measurable tumor volume in the three treatedgroups (p<0.01). The mean tumor volumes in the animals that developedtumors were also affected, 157 mm³ vs 0 mm³, 4 mm³, 34 mm³ (vehicle thendescending dose).

To test the effect of a Compound 1 on established LNCaP tumors, 36castrated SCID mice received AED pellets and were inoculated with LNCaPtumors and monitored as described above. Once the tumors reach 15-25mm³, the mice were paired by tumor volume, and each mouse in a pair wasassigned to the vehicle or 4 mg/mouse/day Compound 1 group. Animals weredosed once a day for three weeks.

The results of this experiment are shown in FIG. 26. Vehicle treatedanimals showed a progressive increase in tumor volume over the course ofthe study. In contrast, treatment with Compound 1 significantly blockedthe growth of tumors (p<0.001). Significant differences in tumor volumesbetween the control and treated groups were observed by the first weekand were maintained through the course of this study (p<0.001). Asignificantly greater percent of mice in the treatment group reducedtheir tumor volumes by 20% or more (p<0.0294). No tumor reduction wasseen in vehicle group. In addition, two mice in the Compound 1 group hada tumor that became non-measurable by Day 15.

Statistical analysis: Time to first measurable tumor volume was analyzedvia Kaplan-Meier product limit estimates, with the exact log-rank testapplied to test for the significance of the difference. Reduction oftumor volume is defined as a reduction in volume of at least 20% of thebaseline volume, persisting to the end of the study. To detect thedifference between active and control group, Fisher's exact test andexact 95% Cl for the difference was applied. A tumor of non-measurablevolume is a tumor that, with the methodology at hand, measures 0 to theend of the study. The growth rate of a tumor was also analyzed via themixed model.

Example 15 Induction of Apoptosis in Cells Undergoing Hyperproliferation

The following study determined the effect of a formulation prepared witha polymorph of 17α-ethynyl-androst-5-ene-3α,17β-diol (crystalline FormIII of Compound 1) on a prostate tumor cell line.

LNCaP Cells (5×10⁵) were seeded in phenol-red free RPMI with 5% hormonedepleted charcoal stripped serum (CSS) in 6-well plates and allowed toadhere overnight, then cultured in fetal bovine serum (FBS) or CSS withor without 50 nM Compound 1 for four days. At the end of the incubationperiod, floating and adherent cells were harvested for cell cycleanalysis. LNCaP cells were resuspended in a 10 mg/mL solution of4,6-diamidino-2-phenylindole (DAPI) and 0.1% NP-40 in a Tris-bufferedsaline solution (pH 7.0) and analyzed using an Influx cytometer(Cytopiea, Seattle, Wash.). Analyses were performed with MultiCyclesoftware (Phoenix Flow Systems, San Diego, Calif.). The Annexin V-FITCApoptosis Detection Kit (Calbiochem, La Jolla, Calif.) was usedaccording to the instructions of the manufacturer to detect apoptosis.

FIG. 27 shows a rise in the number of cells in G1 after incubation ofLNCaP cells with Compound 1. This accumulation of cells in G1 wasaccompanied by an increase in the percentage of apoptotic LNCaP cellsafter exposure to Compound 1 These data suggest is not merely blockingthe growth of LNCaP but acting as a cytotoxic agent for LNCaP cells.

Example 16 Breast Cancer

MNU stimulated tumor model

The following study determined the effect of a formulation prepared witha polymorph of 17α-ethynyl-androst-5-ene-3α,17β-diol (Crystalline FormIII of Compound 1) on the rate of growth and incidence of breast tumorsinduced by the administration of the carcinogen N-methyl-nitrosourea(NMU). Additionally, the activity of Compound 1 was compared toTAMOXIFEN™((Z)-2-[4-(1,2-diphenylbut-1-enyl)phenoxy]-N,N-dimethyl-ethanamine),ARIMIDEX™ (anastrozole) and TAXOTERE™ (docetaxel), which are drugscurrently used to treat breast cancer.

Seven week old female Lewis rats (150 animals, 104 required for thestudy) were anesthetized with isoflurane for NMU administration. NMU wasthe administered ip. at a dose of 50 mg/Kg. Mammary tumors thatdeveloped were measured using a vernier caliper with two axes of thetumor measured in cm. Treatment commenced using a liquid formulation ofCompound 1, prepared by dissolving crystalline Form I in vehicle, whenrats have a tumor volume of 0.5 cm×0.5 cm (at about 12-20 week of age).Treatment continued for 28 consecutive days, followed by 28 days ofobservation. Mammary tumors were removed when a size of 2 cm×2 cm wasreached in accordance with local institutional guidelines.

The experimental groups were Negative control group (no treatment),Vehicle control group (30% cyclodextrin-sulfobutylether in water), twotreatment groups (8 mg/rat and 4 mg/rat), three standard therapy groupsusing an estrogen blocker (TAMOXIFEN™), an aromatase inhibitor(ARIMIDEX™), a cytotoxic agent (TAXOTERE™) and a combination treatmentgroup receiving Compound 1 (8 mg/kg) and TAXOTERE™.

FIG. 28 shows the percent of animals with new tumors plotted againstelapsed time from first day of dosing. This plot shows a longer time toa new tumor as compared to vehicle (p<0.001). Median times to new tumorare 10 days with vehicle compared to at least 56 days with Compound 1,which represents at least a five fold delay to the occurrence of asecond tumor that is attributable to Compound 1. The plot also indicatesthat Compound 1 at 4 mg outperformed TAXOTERE™ (p=0.042) and Compound 1at 8 mg plus TAXOTERE™ is better than TAXOTERE™ alone (p=0.0385).

FIGS. 29 and 30 shows tumor burden by volume for the experimental groupsduring the course of the study. Tumors grow unchecked in the vehicletreated group. Compound 1 treatment consistently shows less tumor burdenthan for the vehicle treated group (Day 7 on: p<0.001). Combinationtreatment with Compound 1 and TAXOTERE™ shows smaller burden profilesthan TAXOTERE™ alone (Day 7 on: p<0.05).

What is claimed is:
 1. A method to make crystalline anhydrate Form III17α-ethynyl-5α-androstane-3α,17β-diol that is characterized by or has(i) an X-ray powder diffraction pattern with peak positions of 9.85,11.33, 15.96, 16.48 and 18.95±0.10 degrees 2-theta or peak positions of15.96, 16.48 and 18.95±0.10 degrees 2-theta and one two or three peakpositions at 9.30, 11.33, 13.45, 16.16 or 17.42±0.10 degrees 2-theta,(ii) an IR-Raman spectroscopy spectrum with peak positions at 1236, 1190and 490 cm⁻¹, optionally with one, two or three peak positions at 1458,1435, 619, 604, 526, 237 or 206 cm⁻¹ or is represented by FIG. 5A orFIG. 5B, and/or (iii) a differential scanning calorimetry thermogramwith a prominent endotherm at 164° C. obtained using a temperature rampof 10° C./min and has a thermogravimetric analysis weight loss of 0.5%or less when heated between 40° C. to 105° C. using a temperature rampof 10° C./min, said method comprising, sufficiently drying at ambienttemperature (about 22-27° C.) under vacuum crystalline solvate Form I17α-ethynyl-5α-androstane-3α,17β-diol, or sufficiently heatingcrystalline solvate Form I 17α-ethynyl-5α-androstane-3α,17β-diol at atemperature(s) from above ambient temperature to a maximum of about 75°C., until the dried or heated material shows a loss on heating of lessthan 0.5% by weight when heated to 50° C.±5°.
 2. The method of claim 1wherein the crystalline solvate Form I17α-ethynyl-5α-androstane-3α,17β-diol starting material is essentiallyfree of the other crystalline or amorphous forms of17α-ethynyl-5α-androstane-3α,17β-diol.
 3. The method of claim 2 whereinthe crystalline solvate Form I 17α-ethynyl-5α-androstane-3α,17β-diolstarting material is at least 95% pure.